Fat-binding polymers

ABSTRACT

The present invention relates to a method for treating obesity, a method for reducing the absorption of dietary fat, and a method for treating hypertriglyceridemia in a patient and to particular polymers for use in the methods or in a manufacture of a medicament. The methods comprise the step of orally administering to a mammal, such as a human, a therapeutically effective amount of one or more fat-binding polymers. The administration of the fat-binding polymer of the invention facilitates the removal of fat from the body prior to digestion, with minimal side effects and low toxicity. In a preferred embodiment, the one or more fat-binding polymers are administered in combination with one or more lipase inibitors, for example, lipstatin and tetrahydrolipstatin.

RELATED APPLICATIONS

[0001] This application is a Continuation of U.S. application Ser. No.09/721,309, filed Nov. 22, 2000, which is a Divisional of U.S.application Ser. No. 09/166,453, filed Oct. 5, 1998, which is aContinuation-in-Part of U.S. application Ser. No. 09/004,963 filed onJan. 9, 1998, the entire content of which is hereby incorporated byreference.

BACKGROUND OF THE INVENTION

[0002] Human obesity is a recognized health problem with approximately97 million people considered clinically overweight in the United States.The accumulation or maintenance of body fat bears a direct relationshipto caloric intake. Therefore, one of the most common methods for weightcontrol to combat obesity is the use of relatively low-fat, low caloriediets, that is, diets containing less fat and calories than a “normaldiet” or that amount generally consumed by the patient.

[0003] The presence of fats in a great many food sources greatly limitsthe food sources which can be used in a low-fat diet. Additionally, fatscontribute to the flavor, appearance and physical characteristics ofmany foodstuffs. As such, the acceptability of low-fat diets and themaintenance of such diets are difficult.

[0004] Various chemical approaches have been proposed for controllingobesity. Anorectic agents, such as dextroamphetamine, the combination ofthe non-amphetamine drugs phentermine and fenfluramine (“Phen-Fen”) anddexfenfluramine (Redux) alone, are associated with serious side effects.Indigestible materials such as OLESTRA™, mineral oil or neopentyl esters(see U.S. Pat. No. 2,962,419) have been proposed as substitutes fordietary fat. Garcinia acid and derivatives thereof have been describedas treating obesity by interfering with fatty acid synthesis. Swellablecrosslinked vinyl pyridine resins have been described as appetitesuppressants via the mechanism of providing non-nutritive bulk, as inU.S. Pat. No. 2,923,662. Surgical techniques, such as temporary ilealbypass surgery, are employed in extreme cases.

[0005] However, methods for treating obesity, such as those describedabove, have serious shortcomings with controlled diet remaining the mostprevalent technique for controlling obesity. As such, new methods fortreating obesity are needed.

SUMMARY OF THE INVENTION

[0006] The present invention relates to a method for treating obesity, amethod for reducing the absorption of dietary fat, and a method fortreating hypertriglyceridemia in a patient and to particular polymersfor use in the methods or in a manufacture of a medicament. The methodscomprise the step of orally administering to a mammal, such as a human,a therapeutically effective amount of a fat-binding polymer. Theadministration of a fat-binding polymer of the invention facilitates theexcretion of fat from the body without digestion, with minimal sideeffects and low toxicity. In a preferred embodiment, the fat-bindingpolymers are administered in combination with a therapeuticallyeffective amount of a lipase inhibitor, such as the pancreatic lipaseinhibitors described in U.S. Pat. No. 4,598,089 to Hadvary et al. Thecombination administration can reduce undesirable side effects oftenencountered when lipase inhibitors, in particular, the pancreatic lipaseinhibitors lipstatin and tetrahydrolipstatin are administered alone. Forexample, a serious side effect resulting from the administration of alipase inhibitor is steatorrhea, or fatty stools.

[0007] The fat-binding polymers of the invention comprise at least onefat-binding region. A fat-binding region can include a region having apositive charge, a region which is hydrophobic or a region having apositive charge and which is hydrophobic.

[0008] In one embodiment, the fat-binding polymer is an aliphaticpolymer selected from the group consisting of polyalkylacrylates,polyacrylamides, polyalkylmethacrylates, polymethacrylamides,poly-N-alkylacrylamides, poly-N-alkylmethacrylamides, substitutedderivatives thereof and copolymers thereof. For example, the substitutedderivatives of the polymers can be characterized by one or moresubstituents, such as substituted or unsubstituted, saturated orunsaturated alkyl, and substituted or unsubstituted aryl groups.Suitable substituents to employ on the alkyl or aryl groups include, butare not limited to, cationic or neutral groups, such as alkoxy, aryl,aryloxy, aralkyl, halogen, amine, and ammonium groups. For example, thepolymer can be poly(dimethylamino propylacrylamide),poly(trimethylammonium ethylacrylate), poly(trimethylammonium ethylmethacrylate), poly(trimethylammonium propyl acrylamide), poly(dodecylacrylate), poly(octadecyl acrylate), poly(octadecyl methacrylate) andcopolymers thereof.

[0009] In another embodiment, the fat binding polymer is a syntheticamine polymer. Amine polymers suitable for use in the invention include,but are not limited to, poly(allylamine), polyethyleneimine,poly(vinylamine), poly(diallylamine), and poly(diallylmethylamine).

[0010] In yet another embodiment, the fat-binding polymer is ahydroxyl-containing polymer, for example, poly(vinylalcohol).

[0011] In a specific embodiment, the fat-binding polymer is anamine-containing polymer wherein one or more hydrophobic regions arebound to a portion of the amine nitrogens of the amine polymer. In aparticular embodiment, between about 1 and about 60 percent of the aminenitrogens are substituted, preferably between about 1 and about 30percent.

[0012] In another embodiment, the hydrophobic region of the fat-bindingpolymer can include a hydrophobic moiety, for example, a substituted orunsubstituted, normal, branched or cyclic alkyl group having at leastfour carbons. In a particular embodiment, the hydrophobic moiety is analkyl group of between about four and thirty carbons.

[0013] In another embodiment, the hydrophobic region is a quaternaryamine-containing moiety having a terminal hydrophobic substituent.Suitable hydrophobic regions which can include a hydrophobic moietyand/or a quaternary amine-containing moiety are described herein and inU.S. Pat. Nos. 5,607,669, 5,679,717 and 5,618,530, the entire contentsof which are incorporated herein by reference in their entirety.

[0014] In yet another embodiment, the fat-binding polymer is substitutedby a lipase inhibitor such as those described in U.S. Ser. No.09/005,379 filed on Jan. 9, 1998, and U.S. Ser. No. 09/166,510 entitled“Lipase Inhibiting Polymers” being filed concurrently herewith, theentire contents of which are incorporated herein by reference.

[0015] The polymers of the present invention offer desirablepharmacological properties such as excellent fat binding properties andlow toxicity. In addition, when the fat-binding polymers areadministered in combination with lipase inhibitors, as described herein,undesirable side effects experienced, such as steatorrhea, when thelipase inhibitors are administered alone can be lessened.

DETAILED DESCRIPTION OF THE INVENTION

[0016] The features and other details of the invention will now be moreparticularly described and pointed out below as well as in the claims.It will be understood that the particular embodiments of the inventionare shown by way of illustration and not as limitations of theinvention. The principle features of this invention can be employed invarious embodiments without departing from the scope of the invention.

[0017] In one aspect, the invention relates to a method for treatingobesity comprising the step of orally administering to a mammal atherapeutically effective amount of one or more fat-binding polymers. Ina preferred embodiment, the fat-binding polymer is administered incombination with a therapeutically effective amount of a lipaseinhibitor.

[0018] In another aspect, the invention relates to a method for reducingthe absorption of dietary fat comprising the step of orallyadministering to a mammal a therapeutically effective amount of one ormore fat-binding polymers. In a preferred embodiment, the fat-bindingpolymer is administered in combination with a therapeutically effectiveamount of a lipase inhibitor.

[0019] In yet another aspect, the invention relates to a method fortreating hypertriglyceridemia in a mammal comprising the step of orallyadministering to a mammal a therapeutically effective amount of one ormore fat-binding polymers. In a preferred embodiment, the fat-bindingpolymer is administered in combination with a therapeutically effectiveamount of a lipase inhibitor.

[0020] A particular aspect of the invention relates to a method fortreating steatorrhea comprising the step of orally administering to amammal a therapeutically effective amount of a fat-binding polymer. In aspecific embodiment, the steatorrhea is a result of the administrationof a lipase inhibitor.

[0021] The invention also relates to fat-binding polymers useful in themethod of the invention.

[0022] “Lipases” as that term is used herein, are ubiquitous enzymeswhich hydrolyze ester bonds in neutral lipids. Examples of lipasesinclude, but are not limited to, pancreatic and gastric lipases. Thepreferred substrates of lipases are insoluble in water. Lipases exhibitmaximal activity in the presence of lipid/water interfaces. For example,pancreatic lipase, which is the key enzyme of dietary triglycerideabsorption, exerts it activity at the water/lipid interface, inconjunction with bile salts and co-lipase.

[0023] “Lipase inhibitor” as that term is used herein refers tocompounds which are capable of inhibiting the action of lipases, forexample, gastric and pancreatic lipases. Lipstatin and its tetrahydroderivative, Tetrahydrolipstatin, as described in U.S. Pat. No. 4,598,089to Hadvary et al., the entire content of which is hereby incorporated byreference, are potent inhibitors of both gastric and pancreatic lipases,as well as cholesterol ester hydrolase. Lipstatin is a natural productof microbial origin, and tetrahydrolipstatin is the result of catalytichydrogenation of lipstatin. Other lipase inhibitors include a class ofcompound commonly referred to as Panclicins. Panclicins are analogues ofTetrahydrolipstatin (See e.g., Mutoh, M., et al., “Panclicins, NovelPancreatic Lipase Inhibitors, II. Structural Elucidation,” The Journalof Antibiotics, 47(12): 1376-1384 (1994), the entire content of which ishereby incorporated by reference.)

[0024] “Fat-binding polymers”, as that term is used herein, are polymerswhich absorb, bind or otherwise associate with fat thereby inhibiting(partially or completely) fat digestion, hydrolysis, or absorption inthe gastrointestinal tract. The fat-binding polymers comprise one ormore fat-binding regions. “Fat-binding regions”, as defined herein caninclude a positively charged region, a hydrophobic region, or a regionwhich is both positively charged and hydrophobic.

[0025] “Fats”, as that term is used herein, are solids or liquid oilsgenerally consisting of glycerol esters of fatty acids. Sources of fatsinclude both animal and vegetable fats, for example, triglyceride estersof saturated and/or unsaturated fatty acids, free fatty acids,diglycerides, monoglycerides, phospholipids and cholesterol esters arefats, as defined herein.

[0026] A variety of polymers can be employed in the invention describedherein. The polymers are synthetic polymers which can be aliphatic, oraromatic. However, aliphatic and synthetic polymers are preferred. A“synthetic polymer”, as that term is employed herein, is a polymer whichis not obtainable from a natural source either directly or throughaminor derivatization of the naturally occurring form. Further, thepolymer can be hydrophobic, hydrophilic or copolymers of hydrophobicand/or hydrophilic monomers. The polymers can be manufactured fromolefinic or ethylenic monomers (such as vinylalcohol, allylamine oracrylic acid) or condensation polymers.

[0027] For example, the polymers can include polyvinylalcohol,polyvinylamine, poly-N-alkylvinylamine, polyallylamine,poly-N-alkylallylamine, polydiallylamine, poly-N-alkyldiallylamine,polyalkylenimine, other polyamines, polyethers, polyamides, polyacrylicacids, polyalkylacrylates, polyacrylamides, polymethacrylic acids,polyalkylmethacrylates, polymethacrylamides, poly-N-alkylacrylamides,poly-N-alkylmethacrylamides, polystyrene, polyvinylnaphthalene,polyethylvinylbenzene, polyaminostyrene, polyvinylbiphenyl,polyvinylanisole, polyvinylimidazolyl, polyvinylpyridinyl,polydimethylaminomethylstyrene, polytrimethylammonium ethylmethacrylate, polytrimethylammonium ethyl acrylate, and substitutedderivatives of the above (e.g., fluorinated monomers thereof) andcopolymers thereof. In addition, the polymers can be furthercharacterized by one or more substituents such as substituents, such assubstituted and unsubstituted, saturated or unsaturated alkyl, andsubstituted or unsubstituted aryl groups. Suitable groups to employinclude cationic or neutral groups, such as alkoxy, aryl, aryloxy,aralkyl, halogen, amine, and ammonium groups.

[0028] Particularly preferred polymers include polyalkylacrylates,polyacrylamides, polyalkylmethacrylates, polymethacrylamides,poly-N-alkylacrylamides, poly-N-alkylmethacrylamides and copolymersthereof. These polymers can be further characterized by one or moresubstituents, such as substituted or unsubstituted, saturated orunsaturated alkyl, and substituted or unsubstituted aryl groups.Suitable substituents include cationic or neutral groups, such asalkoxy, aryl, aryloxy, aralkyl, halogen, amine, and ammonium groups, forexample.

[0029] Other particularly preferred polymers include aliphatic aminepolymers, such as polyallylamine, polydiallylamine,polydiallylmethylamine, polyvinylamine, polyethylenimine. In a specificembodiment, the amine polymer comprises one or more hydrophobic regionswhich are bound to a portion of the amine nitrogens of the aminepolymer. In a particular embodiment, between about 1 and about 60percent of the amine nitrogens are substituted, preferably between about1 and about 30 percent.

[0030] In one embodiment, the hydrophobic region of the fat-bindingpolymer can include a hydrophobic moiety, for example, a substituted orunsubstituted, normal, branched or cyclic alkyl group having at leastfour carbons. In a specific embodiment, the hydrophobic moiety is analkyl group of between about four and thirty carbons.

[0031] In another embodiment, the hydrophobic region is a quaternaryamine-containing moiety having a terminal hydrophobic substituent.

[0032] In yet another embodiment, the fat-binding region comprises anitrogen, for example, the nitrogen of an amine, capable of possessing apositive charge under conditions present in the gastro-intestinal tract.For example, a quaternary amine-containing moiety, or the nitrogen of apolyamine.

[0033] In yet another embodiment, the fat-binding polymer is ahydroxyl-containing polymer, for example, poly(vinylalcohol) which cancomprise further fat-binding regions. For example, the polymer comprisesa repeat unit having the formula

[0034] wherein R is a hydrophobic region.

[0035] Other polymers and methods of preparation, which can be used inthe claimed invention have been reported in the patent literature in,for example, U.S. Pat. Nos. 5,487,888, 5496,545, 5,607,669, 5,618,530,5,624,963, 5,667,775, and 5,679,717 and co-pending U.S. applicationshaving Ser. Nos. 08/471,747, 08/482,969, 08/567,933, 08/659,264,08/823,699, 08/835,857, 08/470,940, 08/461,298, 08/826,197, 08/777,408,08/927,247, 08/964,956, 08/964,498, and 08/964,536, the entire contentsof all of which are incorporated herein by reference.

[0036] The polymer can be linear or crosslinked. Crosslinking can beperformed by reacting the copolymer with one or more crosslinking agentshaving two or more functional groups, such as electrophilic groups,which react with, for example, amine groups to form a covalent bond.Crosslinking in this case can occur, for example, via nucleophilicattack of the polymer amino groups on the electrophilic groups. Thisresults in the formation of a bridging unit which links two or moreamino nitrogen atoms from different polymer strands. Suitablecrosslinking agents of this type include compounds having two or moregroups selected from among acyl chloride, epoxide, and alkyl-X, whereinX is a suitable leaving group, such as a halo, tosyl or mesyl group.Examples of such compounds include, but are not limited to,epichlorohydrin, succinyl dichloride, acryloyl chloride,butanedioldiglycidyl ether, ethanedioldiglycidyl ether, pyromelliticdianhydride, and dihaloalkanes. These crosslinking agents are referredto herein as multifunctional crosslinking agents.

[0037] The polymer composition can also be crosslinked by including amultifunctional co-monomer as the crosslinking agent in thepolymerization reaction mixture. A multifunctional co-monomer can beincorporated into two or more growing polymer chains, therebycrosslinking the chains. Suitable multifunctional co-monomers include,but are not limited to, diacrylates, triacrylates, and tetraacrylates,dimethacrylates, diacrylamides, and dimethacrylamides. Specific examplesinclude ethylene glycol diacrylate, propylene glycol diacrylate,butylene glycol diacrylate, ethylene glycol dimethacrylate, butyleneglycol dimethacrylate, methylene bis(methacrylamide), ethylenebis(acrylamide), ethylene bis(methacrylamide), ethylidenebis(acrylamide), ethylidene bis(methacrylamide), pentaerythritoltetraacrylate, trimethylolpropane triacrylate, bisphenol Adimethacrylate, and bisphenol A diacrylate. Other suitablemultifunctional monomers include polyvinylarenes, such asdivinylbenzene.

[0038] The amount of cross-linking agent is typically between about 0.5and about 25 weight % based on the combined weight of crosslinking agentand monomers, with 1-20% being preferred. Typically, the amount ofcross-linking agent that is reacted with the polymer, when thecrosslinking agent is a multifunctional agent, is sufficient to causebetween about 0.1 and 20 percent of the nucleophiles present on themonomer, for example, an amine to react with the crosslinking agent. Ina preferred embodiment, between about 3 and 15 percent of thenucleophilic sites, for example, amines react with the multifunctionalcrosslinking agent.

[0039] The hydrophobic region or regions of the fat-binding polymersinclude but are not limited to, for example, a hydrophobic moiety suchas a substituted or unsubstituted, normal, branched or cyclic alkylgroup having at least about four carbons. For example, a hydrophobicmoiety such as an alkyl group of at least four carbons can be bound tothe fat-binding polymer, for example, through an amine of thefat-binding polymer.

[0040] A “hydrophobic moiety”, as the term is used herein, is a moietywhich, as a separate entity, is more soluble in octanol than water. Forexample, the octyl group (C₈H₁₇) is hydrophobic because its parentalkane, octane, has greater solubility in octanol than in water. Thehydrophobic moieties can be a saturated or unsaturated, substituted orunsubstituted hydrocarbon group. Such groups include substituted andunsubstituted, normal, branched or cyclic alkyl groups having at leastfour carbon atoms, substituted or unsubstituted arylalkyl orheteroarylalkyl groups and substituted or unsubstituted aryl orheteroaryl groups. Preferably, the hydrophobic moiety includes an alkylgroup of between about four and thirty carbons. Specific examples ofsuitable hydrophobic moieties include the following alkyl groupsn-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, n-undecyl, n-dodecyl,n-tetradecyl, n-octadecyl, 2-ethylhexyl, 3-propyl-6-methyl decyl andcombinations thereof. Other examples of suitable hydrophobic moietiesinclude haloalkyl groups of at least six carbons (e.g., 10-halodecyl),hydroxyalkyl groups of at least six carbons (e.g., 11-hydroxyundecyl),and aralkyl groups (e.g., benzyl).

[0041] The positively charged region or regions of the fat-bindingpolymers include but are not limited to, for example, an amine nitrogencapable of possessing a positive charge under conditions present in thegastro-intestinal tract and a quaternary amine-containing moiety.Suitable quaternary amine-containing moieties include alkyltrialkylammonium groups also referred to as ammonioalkyl groups. Theterm, “ammonioalkyl”, as used herein, refers to an alkyl group which issubstituted by a nitrogen bearing three additional substituents. Thus,the nitrogen atom is an ammonium nitrogen atom which bears an alkylenesubstituent, which links the ammonium nitrogen atom to the polymer, andthree additional terminal alkyl substituents having from about one toabout twenty-four carbons. A “terminal substituent” of the quaternaryamine-containing moiety, as the term is employed herein, is any one ofthe three substituents on the quaternary amine nitrogen which is not thecarbon chain between the polymer backbone and the nitrogen of thequaternary ammonium center. In a specific embodiment, the polymer is anamine polymer and the alkylene group links the ammonium nitrogen atom tothe nitrogen atom of the polymer. It is to be understood that multiplemoieties can be bound to the same amine and/or different amines of thepolymer composition.

[0042] In another embodiment, the quaternary amine-containing moiety canbear at least one terminal hydrophobic alkyl substituent, such as analkyl group having between about four and twenty-four carbons, therebyproviding both a hydrophobic region and a positively charged region incombination.

[0043] An ammonioalkyl group will further include a negatively chargedcounterion, such as a conjugate base of a pharmaceutically acceptableacid. Examples of suitable counterions include Cl⁻, PO₄ ⁻, Br⁻, CH₃SO₃⁻, HSO₄ ⁻, SO₄ ²⁻, HCO₃ ⁻, CO₃ ²⁻, acetate, lactate, succinate,propionate, butyrate, ascorbate, citrate, maleate, folate, an amino acidderivative, and a nucleotide.

[0044] Suitable ammonioalkyl groups are of the general formula:

[0045] wherein, R¹, R² and R³ represent an alkyl group, wherein eachR¹-R³, independently, is a normal or branched, substituted orunsubstituted alkyl group having a carbon atom chain length of betweenabout one to about twenty-four carbon atoms, n is an integer having avalue of two or more and Y is a negatively charged counterion. In aparticular embodiment, R¹, R² and R³ are all methyl groups and n is aninteger between about 2 and about 12 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10,11, or 12).

[0046] The alkyl group, which provides the alkylene linking groupbetween the polymer, for example, and the amine of the amine-containingmonomer or repeat unit, and the ammonium nitrogen of the alkyltrialkylammonium group, is two or more carbon atoms in length. Examplesof preferred alkylene linking groups are ethyl, propyl, butyl, pentyl,hexyl, octyl, and decyl groups. Example of suitable quaternaryamine-containing moieties include, but are not limited to:

[0047] 3-(trimethylammonio)propyl;

[0048] 4-(trimethylammonio)butyl;

[0049] 6-(trimethylammonio)hexyl;

[0050] 8-(trimethylammonio)octyl;

[0051] 10-(trimethylammonio)decyl;

[0052] 12-(trimethylammonio)dodecyl and combinations thereof Aparticularly preferred amine-containing moiety is a6-(trimethylammonio)hexyl group.

[0053] Alternatively, a quaternary amine-containing moiety and ahydrophobic moiety are present in the same substituent, therebyproviding both a positively charged and hydrophobic region incombination. For example, the quaternary amine nitrogen or ammoniumnitrogen of the quaternary amine-containing moiety is bound to thepolymer backbone by an alkylene having two or more carbons. However, atleast one of the three terminal substituents (R¹, R² and R³) of theammonium nitrogen is a hydrophobic alkyl group having from four to abouttwenty-four carbons. The remaining terminal substituents are eachindependently a normal or branched, substituted or unsubstituted alkylgroup having from one to about twenty-four carbons or a hydrogen atom.In another embodiment, at least two of the three terminal substituentscan be hydrophobic alkyl groups having from four to about twenty-fourcarbons, the remainder having from one to about twenty-four carbons or ahydrogen atom. In a further embodiment, all three of the terminalsubstituents can be hydrophobic alkyl groups having from six to abouttwenty-four carbons.

[0054] A “hydrophobic alkyl group”, as that term is employed herein,includes a substituted or unsubstituted alkyl group having from four toabout twenty-four carbons and which is hydrophobic, as earlier defined.The hydrophobic alkyl group can be, for example, a normal or branched,substituted or unsubstituted alkyl group having from six to abouttwenty-four carbons.

[0055] Particular examples of quaternary amine-containing moieties,which provide both a hydrophobic and quaternary amine-containingsubstituent, include, but are not limited to:

[0056] 4-(dioctylmethylammonio)butyl;

[0057] 3-(dodecyldimethylammonio)propyl;

[0058] 3-(octyldimethylammonio)propyl;

[0059] 3-(decyldimethylammonio)propyl;

[0060] 5-(dodecyldimethylammonio)pentyl;

[0061] 6-(dimethyldecylammonio)hexyl;

[0062] 6-(decyldimethylammonio)hexyl;

[0063] 3-(tridecylammonio)propyl;

[0064] 3-(docosyldimethylammonio)propyl;

[0065] 6-(docosyldimethylammonio)hexyl;

[0066] 4-(dodecyldimethylammonio)butyl;

[0067] 3-(octadecyldimethylammonio)propyl;

[0068] 3-(hexyldimethylammonio)propyl;

[0069] 3-(methyldioctylammonio)propyl;

[0070] 3-(didecylmethylammonio)propyl;

[0071] 3-(heptyldimethylammonio)propyl;

[0072] 3-(dimethylnonylammonio)propyl;

[0073] 6-(dimethylundecylammonio)hexyl;

[0074] 4-(heptyldimethylammonio)butyl;

[0075] 4-(dioctylmethylammonio)butyl;

[0076] 6-(octyldimethylammonio)hexyl;

[0077] 12-(decyldimethylammonio)dodecyl;

[0078] 3-(dimethylundecylammnio)propyl; and

[0079] 3-(tetradecyldimethylammonio)propyl.

[0080] Other suitable quaternary amine-containing moieties includesecondary and tertiary analogs, such as4-(dioctylmethylammonio)₄-methylbutyl and4-(dioctylmethylammonio)-4,4-dimethylbutyl.

[0081] The fat-binding polymers of the invention can be formed, forexample, by reacting a polymer, which can be linear or crosslinked, witha suitable alkylating agent or by polymerizing an alkylated monomer.

[0082] An “acylating agent”, as that term is employed herein, means areactant that, when reacted with a monomer or a copolymer characterizedby a repeat unit of the invention and having a nucleophilic site capableof reaction with the acylating agent, causes an acyl substituent, inparticular a hydrophobic acyl substituent, as described herein, to becovalently bound to one or more of sites on the fat-binding polymer, forexample, the amine nitrogen atoms or hydroxyl oxygens of anamine-containing or hydroxyl-containing monomer or polymer,respectively. Further, when multiple substituents are employed, they canbe bound to the same and/or different nucleophilic sites of thefat-binding polymer, for example, the same and/or different aminenitrogens of an amine-containing fat-binding polymer or hydroxyl oxygenof a hydroxyl-containing polymer.

[0083] Suitable acylating agents are compounds comprising an acyl groupor acyl derivative, for example an anhydride. For example, when theacylating agent is acetic anhydride the nucleophile is modified byaddition of an acetyl group. Acylating agents suitable for the additionof a hydrophobic moiety contain an acyl group having at least fourcarbon atoms, which is bonded to a leaving group such as a halo (e.g.,chloro, bromo or iodo). Activated esters are also suitable acylatingagents. Examples of suitable acylating agent which provide a hydrophobicmoiety include acyl halides having at least four carbon atoms, such asbutyryl halide, valeryl halide, hexanoyl halide, heptanoyl halide,octanoyl halide, nonanoyl halide, decanoyl halide, undecanoyl halide,and combinations thereof.

[0084] An “alkylating agent”, as that term is employed herein, means areactant that, when reacted with a monomer or a copolymer characterizedby a repeat unit of the invention and having a nucleophilic site capableof reaction with the alkylating agent, causes a hydrophobic substituent,as described herein, to be covalently bound to one or more of sites onthe fat-binding polymer, for example, the amine nitrogen atoms orhydroxyl oxygens of an amine-containing or hydroxyl-containing monomeror polymer, respectively. Further, when multiple substituents areemployed, they can be bound to the same and/or different nucleophilicsites of the fat-binding polymer, for example, the same and/or differentamine nitrogens of an amine-containing fat-binding polymer or hydroxyloxygen of a hydroxyl-containing polymer.

[0085] Suitable alkylating agents are compounds comprising an alkylgroup or alkyl derivative, having at least four carbon atoms, which isbonded to a leaving group such as a halo (e.g., chloro, bromo or iodo),tosylate, mesylate or epoxy group).

[0086] Examples of suitable alkylating agents which provide ahydrophobic moiety include alkyl halides having at least four carbonatoms, such as n-hexyl halide, n-heptyl halide, n-octyl halide, n-nonylhalide, n-decyl halide, n-undecyl halide, n-dodecyl halide, n-tetradecylhalide, n-octadecyl halide, and combinations thereof. Other examplesinclude: a dihaloalkane that includes an alkyl group of at least fourcarbons (e.g., a 1,10-dihalodecane); a hydroxyalkyl halide having atleast four carbon atoms (e.g., an 11-halo-1-undecanol); an aralkylhalide (e.g., a benzyl halide); an alkyl epoxy ammonium salt having atleast six carbons (e.g., glycidylpropyl-trimethylammonium salts) andepoxyalkylamides having at least six carbons (e.g., N-(2,3-epoxypropyl)butyramide or N-(2,3-epoxypropyl) hexanamide). Preferred halogencomponents of the alkyl halides are bromine and chlorine. Particularlypreferred alkylating agents which, when reacted with the polymercomposition, will cause formation of an amine polymer reaction productthat includes a first substituent, are 1-bromodecane and 1-chlorooctane.

[0087] Examples of suitable alkylating agents which can provide aquaternary amine-containing moiety have the general formula:

[0088] wherein,

[0089] R¹, R², and R³ represent an alkyl group, wherein each Rindependently is a normal or branched, substituted or unsubstitutedalkyl group having a carbon atom chain length of between about one toabout twenty four carbon atoms,

[0090] n is an integer having a value of two or more,

[0091] X is a leaving group as earlier described, and

[0092] Y is a negatively charged counterion.

[0093] When at least one of the three terminal substituents of thequaternary amine alkylating agent is a hydrophobic alkyl group havingfrom four to about twenty-four carbons, the alkylating agent thereforeprovides both a hydrophobic moiety and a quaternary amine-containingmoiety. The alkylene group in this instance is three or more carbonatoms in length.

[0094] Particular examples of quaternary ammonium compounds suitable asalkylating agents include the following:

[0095] (4-bromobutyl)dioctylmethylammonium bromide;

[0096] (3-bromopropyl)dodecyldimethylammonium bromide;

[0097] (3-chloropropyl)dodecyldimethylammonium bromide;

[0098] (3-chloropropyl)decyldimethylammonium bromide;

[0099] (5-tosylpentyl)dodecyldimethylammonium bromide;

[0100] (6-bromohexyl)dimethyldecylammonium bromide;

[0101] (12-bromododecyl)decyldimethylammonium bromide;

[0102] (3-bromopropyl)tridecylammonium bromide;

[0103] (3-bromopropyl)docosyldimethylammonium bromide;

[0104] (6-bromohexyl)docosyldimethylammonium bromide;

[0105] (4-chlorobutyl)dodecyldimethylammonium bromide;

[0106] (3-chloropropyl)octadecyldimethylammonium bromide;

[0107] (3-bromopropyl)octyldimethylammonium bromide;

[0108] (4-iodobutyl)dioctylmethylammonium bromide;

[0109] (2,3-epoxy propyl)decyldimethylammonium bromide; and

[0110] (6-bromohexyl)docosyldimethyammonium bromide.

[0111] Other suitable alkylating agents include secondary and tertiaryanalogs, such as (3-bromobutyl)dioctylmethylammonium bromide and(3-chloro-3,3-dimethyl propyl)dioctylmethylammonium bromide.

[0112] Examples of suitable alkyl trimethylammonium alkylating agentsinclude alkyl halide trimethylammonium salts, such as:

[0113] (4-halobutyl)trimethylammonium salt;

[0114] (5-halopentyl)trimethylammonium salt;

[0115] (6-halohexyl)trimethylammonium salt;

[0116] (7-haloheptyl)trimethylammonium salt;

[0117] (8-halooctyl)trimethylammonium salt;

[0118] (9-halononyl)trimethylammonium salt;

[0119] (10-halodecyl) trimethylammonium salt;

[0120] (11-haloundecyl)trimethylammonium salt;

[0121] (12-balododecyl)trimethylammonium salt; and combinations thereof.A particularly preferred quaternary amine-containing alkylating agent is(6-bromohexyl)-trimethylammonium bromide.

[0122] In another embodiment, the fat-binding polymer can be have alipase inhibitor covalently bound to the polymer as described in U.S.Ser. No. 09/005,379 filed on Jan. 9, 1998, and U.S. Ser. No. 09/166,510,entitled “Lipase Inhibiting Polymers” being filed concurrently herewith,the entire contents of both of which are incorporated herein byreference. In a further embodiment, the fat-binding polymer can beadministered in combination with a lipase inhibitor which is convalentlybound to a polymer as described in U.S. Ser. No. 09/005,379 filed onJan. 9, 1998, and U.S. Ser. No. 09/166,510 entitled “Lipase InhibitingPolymers” being filed concurrently herewith, the entire contents ofwhich are incorporated herein by reference.

[0123] As used herein, the terms “therapeutically effective amount” and“therapeutic amount” are synonymous. The terms refer to an amount whichis sufficient to treat obesity, reduce the absorption of fat or treathypertriglyceridemia. The dosage of fat-binding polymer administered tothe patient will vary depending among other things on the weight of thepatient and the general health of the patient. The dosage can bedetermined with regard to established medical practice. The amount offat-binding polymer administered can be in the range of from about 0.01mg/kg of body weight/day to about 1 g/kg of body weight/day. The amountof lipase inhibitor which can be administered in combination with thefat-binding polymers of the invention can be determined with regard toaccepted medical practice.

[0124] As disclosed above, in a preferred embodiment, the fat-bindingpolymer is administered in combination with a lipase inhibitor, asdescribed herein. The term “in combination” in this context includesboth simultaneous or sequential administration (either type of compoundfirst) of the fat-binding polymer and lipase inhibitor. The fat-bindingpolymer and lipase inhibitor, when used in combination, can be employedtogether in the same dosage form or in separate dosage forms taken atthe same time or within a time period, wherein both the fat-bindingpolymer and lipase inhibitor are present in a therapeutically effectiveamount.

[0125] The fat-binding polymers of the invention can be formulated usingconventional inert pharmaceutical adjuvant materials into dosage formswhich are suitable for oral administration. The oral dosage formsinclude tablets, capsules, suspension, solutions, and the like. Theidentity of the inert adjuvant materials which are used in formulatingthe fat-binding polymers of the invention will be immediately apparentto persons skilled in the art. These adjuvant materials, eitherinorganic or organic in nature, include, for example, gelatin, albumin,lactose, starch, magnesium stearate, preservatives (stabilizers),melting agents, emulsifying agents, salts, and buffers.

[0126] In patients with hypertriglyceridemia it is to be understood thatthe patient does not necessarily suffer from hypercholesterolemia.

EXEMPLIFICATIONS EXAMPLE 1 Synthesis of Diallylamine-HCl (DAA-HCl)Solution

[0127] Diallylamine (DAA) (2000.3 g) was added slowly over a period of 2days to concentrated HCl (2035.6 g). The temperature of the reaction wasmaintained below 10° C. by cooling the flask in an ice-salt-water bath,and by adjusting the addition rate. The room temperature pH of theresulting DAA-HCl solution (68.16% DAA-HCl) was 0.005.

EXAMPLE 2 Polymerization of Diallylamine-HCl

[0128] To a 12-liter, 4-necked, round-bottomed flask equipped with anoverhead stirrer and an air condenser was added DAA-HCl (3667.8 g of a68.16% solution, and deionized water (4665.5 g). The resulting solutionhad a pH of 0.741. To the flask was added NaOH (66.8 g of a 50% aqueoussolution). The resulting solution had a pH of 2.554. Nitrogen gas wasbubbled through the solution, via a stainless steel needle, withstirring, and venting on top of the air condenser for 2 hours. Thenitrogen line was put on top of the air condenser with positive pressurefrom a mineral oil bubbler. To the flask was added 125.0 g of freshlymade 20% V-50 (Wako Chemicals USA, Inc., Richmond, Va.) in deionizedwater. This was added via syringe through a septum. The V-50 solutionwas not degassed with nitrogen. The solution was heated to 60° C. over aperiod of 1 hour and 8 minutes, with a heating mantle connected to aJ-Kem temperature controller. The solution was heated at 60° C. for 18hours. After the first 18 hour heating period, the reaction solution wasallowed to cool down slowly to 49° C., and to the flask was added 125.0g of freshly made 20% V-50 in deionized water. The solution was heatedto 60° C. over a period of about 15 minutes, with a heating mantleconnected to a J-Kem temperature controller. The solution was heated at60° C. for 18 hours. After the second 18 hour heating period, thereaction solution was allowed to cool down slowly to 40° C., and to theflask was added 125.0 g of freshly made 20% V-50 in deionized water. Thesolution was heated to 60° C. over a period of about 15 minutes, with aheating mantle connected to a J-Kem temperature controller. The solutionwas heated at 60° C. for 18 hours. After cooling to room temperature,the solution was a dark orange viscous, flowable, clear solution. Theflask contents were combined with deionized water (4166.7 g). Theresulting solution had a pH of 4.4. SEC analysis: Mw 61494 Daltons;Polydispersity 2.43.

EXAMPLE 3 Crosslinking of a Soluble Polymer to Obtain a Insoluble Gel;Preparation of 3 Mol % Crosslinked Poly(Allylamine)HCl

[0129] Poly(allylamine) Hydrochloride (200 g of 50% aqueous solution,1.07 mol monomer equivalents) was dissolved in a mixture of ethanol (213mL) and water (125 mL) in a 1-liter, round-bottomed flask equipped withan overhead mechanical stirrer. The pH of the solution was brought to10.0-10.2 by the addition of NaOH (50% solution). Epichlorohydrin (2.97g, 32.07 mmol) was then added to the rapidly stirred solution in oneportion. This mixture was stirred at room temperature (19-22° C.) untila gel formed (approx. 30 min), then stirring was suspended and themixture was allowed to sit at room temperature for 20 hours. After the20 hour reaction time had elapsed, the gel was transferred into a5-liter bucket with 3 liters of deionized water. The mixture was thenstirred with an overhead mechanical stirrer until the gel was welldispersed in solution. The pH was then adjusted to <1 using concentratedHCl. The mixture was then vacuum filtered through Whatman 541 filterpaper. The filtered polymer gel was then collected and purified bysuspension into 4 liters of deionized water followed by vacuumfiltration through Whatman 541 filter paper. The procedure of suspensioninto deionized water followed by vacuum filtration was repeated severaltimes until the conductivity of the suspended polymer gel was <0.5mS/cm. After the final vacuum filtration, the polymer gel wastransferred into several Pyrex drying trays and placed into a convectionoven at 70° C. to dry (24-48 hours). The dried solid was ground to afine powder using a lab mill with stainless steel blades, and was passedthrough a sieve (50 mesh) to remove large granules. The ground productwas then placed in a vacuum oven at 60° C. and 28 mm Hg for at least 16hours. Yield=83%. TABLE 1 Epichlorohydrin Crosslinking Reactions Using aProcedure Similar to Example 3 Example Polyamine mol % Xlink 4Poly(allylamine) HCl 6 5 Poly(allylamine) HCl 9 6 Poly(allylamine) HCl10 mol % C 12H 25 3 (prepared according to Example 82) 7Polyethylenimine 3 8 Polyethylenimine 6 9 Poly(diallylamine) HCl 3 10 Poly(diallylamine) HCl 4.5 11  Poly(diallylamine) HCl 6 12 Poly(diallylmethylamine) HCl 4.5 13  Poly(vinylamine) 4.5

EXAMPLE 14 Low Level Crosslinking of a Soluble Polymer to Obtain a HighMolecular Weight Soluble Polymer; Preparation of 0.75 Mol % CrosslinkedSoluble Poly(Allylamine)HCl

[0130] Poly(allylamine) Hydrochloride (200 g of 50% aqueous solution,1.07 mol monomer equivalents) was dissolved a mixture of ethanol (213mL) and water (125 mL) in a 1-liter, round-bottomed flask equipped withan overhead mechanical stirrer. The pH of the solution was brought to10.0-10.2 by the addition of NaOH (50% solution). Epichlorohydrin (743mg, 8.03 mmol) was then added to the rapidly stirred solution in oneportion at room temperature (19-22° C.). This mixture was stirred atroom temperature (19-22° C.) for 20 hours. After the 20 hour reactiontime had elapsed, the pH was adjusted to 11.5-12.0 by the addition of a50% NaOH solution. The reaction mixture was then poured into a 5-literbeaker containing 2 liters of methanol stirred with an overheadmechanical stirrer. A fine precipitate was observed as this mixture wasstirred for 30 minutes. The mixture was vacuum filtered through Whatman541 filter paper, and the clear filtrate was acidified with concentratedHCl (pH<1) producing a thick polymer precipitate and a cloudy solution.The cloudy methanol solution was decanted away from the crude solidproduct. The precipitate was dissolved in a minimum amount of water(approx. 300 mL) and acidified with concentrated HCl to a pH of <2. Theaqueous polymer solution was then poured with overhead mechanicalstirring into a 3-liter beaker containing at least 5 volumes (approx.1.5 liters) of methanol (Isopropanol can be used in place of methanol inthis step). The polymeric product precipitated as a white solid. Afterstirring for 15 minutes, the precipitate was separated from solution bydecantation and suspended in 2 liters of isopropyl alcohol. The solidwas broken up using a metal spatula and the mixture was stirred for 2hours. The isopropyl alcohol was then removed by decanting and theproduct was again suspended in 2 liters of fresh isopropyl alcohol.After 2 hours of stirring, the solvent was decanted away and the solidproduct was placed in a convection oven at 70° C. to dry (24-48 hours).The dried solid was ground to a fine powder using a lab mill withstainless steel blades, and was passed through a sieve (50 mesh) toremove large granules. The ground product was then placed in a vacuumoven at 60° C. and 28 mmHg for at least 16 hours. Yield=88%.

EXAMPLE 15 Preparation of 0.75 Mol % Crosslinked Poly(Diallylamine)HCl

[0131] Poly(diallylamine) Hydrochloride (3250 g of 20% aqueous solution,4.86 mol monomer equivalents) was placed in a 20-liter bucket equippedwith an overhead mechanical stirrer. The pH of the solution was broughtto 10.6 by the addition of NaOH (50% solution). Epichlorohydrin (2.86mL, 0.037 mol) was then added to the rapidly stirred solution in oneportion at room temperature (19-22° C.). This mixture was stirred atroom temperature (19-22° C.) for 20 hours. A viscous solution resulted.Methanol (10 liters) was added, and the pH was adjusted to >11.5 using a50% NaOH solution. This solution was then filtered to remove insolublecrosslinked polymer. The clear filtrate was acidified with concentratedHCl to a pH of <2, and the polymer product was precipitated with theaddition of a large volume of ethanol. The solid was collected bydecantation and washed with isopropanol. The solid product was thenplaced in a convection oven at 70° C. to dry (24-48 hours). The driedsolid was ground to a fine powder using a lab mill with stainless steelblades, and was passed through a sieve (50 mesh) to remove largegranules. The ground product was then placed in a vacuum oven at 60° C.and 28 mmHg for at least 16 hours. Yield=355 g

EXAMPLE 16 Alkylation of an Insoluble Gel Preparation of 10 Mol %Dodecyl Substituted, 3 Mol % Crosslinked Poly(Allylamine)HCl

[0132] Epichlorohydrin (3 mol %) crosslinked poly(allylamine)HCl (100 gof dry solid, 1.05 mol monomer equivalents) was suspended in methanol(1250 mL) in a 3-liter, round-bottomed flask equipped with an overheadmechanical stirrer, a condenser, and a thermocouple probe. Deionizedwater (750 mL) was slowly added to the suspension with good stirring,and the mixture was stirred until a uniform suspension resulted (approx.3 hours). The mixture was then heated to 70° C., and the pH of thesolution was brought to 10.0-10.2 by the addition of NaOH (50%solution). 1-Bromododecane (26.17 g, 0.105 mol) was then added to thestirred solution in one portion. This mixture was stirred at 70° C. for20 hours. The solution pH was checked periodically during this time, andwas maintained at 10.0-10.2 by the addition of small quantities of 50%NaOH. After the 20 hour reaction time had elapsed, the mixture wascooled to room temperature and the pH was adjusted to <1 usingconcentrated HCl. The reaction mixture was then poured into a 5-literbeaker containing 3 liters of methanol stirred with an overheadmechanical stirrer. The mixture was stirred until a uniform suspensionresulted. The mixture was then vacuum filtered through Whatman 541filter paper. The filtered polymer gel was collected and suspended in 3liters of fresh methanol. The methanol suspension was acidified to a pHof <1 with concentrated HCl. The mixture was then vacuum filteredthrough Whatman 541 filter paper. The filtered polymer gel was collectedand suspended in 4-liters of 2M aqueous NaCl. The aqueous suspension wasacidified to a pH of <1 with concentrated HCl. The mixture was thenvacuum filtered through Whatman 541 filter paper. The filtered polymergel was then collected and purified by suspension into 4 liters ofdeionized water followed by vacuum filtration through Whatman 541 filterpaper. The procedure of suspension into deionized water followed byvacuum filtration was repeated several times until the conductivity ofthe suspended polymer gel was <0.5 mS/cm. After the final vacuumfiltration, the polymer gel was transferred into several Pyrex dryingtrays and placed into a convection oven at 70° C. to dry (24-48 hours).The dried solid was ground to a fine powder using a lab mill withstainless steel blades, and was passed through a sieve (50 mesh) toremove large granules. The ground product was then placed in a vacuumoven at 60° C. and 28 mmHg for at least 16 hours. Yield=82%. TABLE 2Alkylation Reactions of Xlinked Polyamines Using the Procedure inExample 16 Example Polyamine Alkylating Agent mol % 17Poly(allylamine)HCl Xlink 3 mol % 1-Bromohexane 5 18 Poly(allylamine)HClXlink 3 mol % 1-Bromohexane 10 19 Poly(allylamine)HCl Xlink 3 mol %1-Bromohexane 25 20 Poly(allylamine)HCl Xlink 3 mol % 1-Bromohexane 5021 Poly(allylamine)HCl Xlink 3 mol % 1-Bromooctane 5 22Poly(allylamine)HCl Xlink 3 mol % 1-Bromooctane 10 23Poly(allylamine)HCl Xlink 3 mol % 1-Bromooctane 25 24Poly(allylamine)HCl Xlink 3 mol % 1-Bromooctane 50 25Poly(allylamine)HCl Xlink 3 mol % 1-Bromododecane 5 26Poly(allylamine)HCl Xlink 3 mol % 1-Bromododecane 25 27Poly(allylamine)HCl Xlink 3 mol % 1-Bromododecane 50 28Poly(allylamine)HCl Xlink 3 mol % 1-Bromooctadecane 5 29Poly(allylamine)HCl Xlink 3 mol % 1-Bromooctadecane 10 30Poly(allylamine)HCl Xlink 3 mol % 1-Bromooctadecane 25 31Poly(allylamine)HCl Xlink 3 mol % 1-Bromo-2-ethylhexane 5 32Poly(allylamine)HCl Xlink 3 mol % 1-Bromo-2-ethylhexane 10 33Poly(allylamine)HCl Xlink 3 mol % 1-Bromo-2-ethylhexane 25 34Poly(allylamine)HCl Xlink 3 mol % 1-Bromo-2-ethylhexane 50 35Poly(allylamine)HCl Xlink 3 mol % (Bromomethyl)cyclohexane 5 36Poly(allylamine)HCl Xlink 3 mol % (Bromomethyl)cyclohexane 10 37Poly(allylamine)HCl Xlink 3 mol % (Bromomethyl)cyclohexane 25 38Poly(allylamine)HCl Xlink 3 mol % (3-Bromopropyl)trimethyl- 5 ammoniumBromide 39 Poly(allylamine)HCl Xlink 3 mol % (3-Bromopropyl)trimethyl-10 ammonium Bromide 40 Poly(allylamine)HCl Xlink 3 mol %(3-Bromopropyl)trimethyl- 25 ammonium Bromide 41 Poly(allylamine)HClXlink 3 mol % (6-Bromohexyl)trimethyl- 10 ammonium Chloride 42Poly(allylamine)HCl Xlink 3 mol % 1,3-Propane Sultone 5 43Poly(allylamine)HCl Xlink 3 mol % 1,3-Propane Sultone 10 44Poly(allylamine)HCl Xlink 3 mol % 1,3-Propane Sultone 25 45Poly(allylamine)HCl Xlink 3 mol % 1-Bromoacetic Acid 5 46Poly(allylamine)HCl Xlink 3 mol % 1-Bromoacetic Acid 10 47Poly(allylamine)HCl Xlink 3 mol % 1-Bromoacetic Acid 25 48Poly(allylamine)HCl Xlink 3 mol % 2-Bromoethanesulfonic Acid 5 Na salt49 Poly(allylamine)HCl Xlink 3 mol % 2-Bromoethanesulfonic Acid 10 Nasalt 50 Poly(allylamine)HCl Xlink 3 mol % 2-Bromoethanesulfonic Acid 25Na salt 51 Poly(allylamine)HCl Xlink 3 mol % 2-Bromododecane 10(6-Bromohexyl) 10 trimethylammonium Chloride 52 Poly(allylamine)HClXlink 6 mol % 2-Bromododecane 10 53 Poly(allylamine)HCl Xlink 6 mol %2-Bromododecane 10 (6-Bromohexyl) 10 trimethylammonium Chloride 54Poly(allylamine)HCl Xlink 3 mol % (4-chlorobutyl) 10dodecyldimethylammonium Bromide 55 Poly(allylamine)HCl Xlink 3 mol %(4-chlorobutyl) 40 dodecyldimethylammonium Bromide 56 Poly(ethylenimine)Xlink 3 mol % 1-Bromohexane 5 57 Poly(ethylenimine) Xlink 3 mol %1-Bromohexane 10 58 Poly(ethylenimine) Xlink 3 mol % 1-Bromohexane 25 59Poly(ethylenimine) Xlink 3 mol % 1-Bromooctane 5 60 Poly(ethylenimine)Xlink 3 mol % 1-Bromooctane 10 61 Poly(ethylenimine) Xlink 3 mol %1-Bromooctane 25 62 Poly(ethylenimine) Xlink 3 mol % 1-Bromododecane 563 Poly(ethylenimine) Xlink 3 mol % 1-Bromododecane 10 64Poly(ethylenimine) Xlink 3 mol % 1-Bromododecane 25 65Poly(diallylamine)HCl Xlink 4.5 1-Bromohexane 5 mol % 66Poly(diallylamine)HCl Xlink 4.5 1-Bromohexane 25 mol % 67Poly(diallylamine)HCl Xlink 4.5 1-Bromohexane 50 mol % 68Poly(diallylamine)HCl Xlink 4.5 1-Bromooctane 5 mol % 69Poly(diallylamine)HCl Xlink 4.5 1-Bromooctane 30 mol % 70Poly(diallylamine)HCl Xlink 4.5 1-Bromooctane 40 mol % 71Poly(diallylamine)HCl Xlink 4.5 1-Bromododecane 5 mol % 72Poly(diallylamine)HCl Xlink 4.5 1-Bromododecane 11 mol % 73Poly(diallylamine)HCl Xlink 4.5 1-Bromododecane 25 mol % 74Poly(diallylamine)HCl Xlink 4.5 (4-chlorobutyl) 10 mol %dodecyldimethylammonium Bromide 75 Poly(diallylamine)HCl Xlink 4.5(4-chlorobutyl) 20 mol % dodecyldimethylammonium Bromide 76Poly(diallylamine)HCl Xlink 4.5 (4-chlorobutyl) 30 mol %dodecyldimethylammonium Bromide

EXAMPLE 77 Acetylation of a Crosslinked Polymer Gel Preparation of 25Mol % Acetylated-Poly(Allylamine)HCl

[0133] Epichlorohydrin (3 mol %) crosslinked poly(allylamine)HCl (57.4 gof dry solid, 0.602 mol monomer equivalents) was suspended in methanol(1 liter) in a 2-liter, round-bottomed flask equipped with an overheadmechanical stirrer, a condenser, and a thermocouple probe. Deionizedwater (550 mL) was slowly added to the suspension with good stirring,and the mixture was stirred until a uniform suspension resulted (approx.3 hours). The mixture was then cooled to 15° C. with an ice bath, andthe pH of the solution was brought to 9.5 by the addition of NaOH (50%solution). Acetic anhydride (15.41 g, 0.151 mol) was then added to thestirred solution in one portion. This mixture was stirred at 15° C. for30 minutes. The solution pH was maintained at 9.5 during this time bythe addition of small quantities of 50% NaOH. After the 30 minutes itwas observed that the pH of the mixture was stable. The crude polymerproduct was then purified by suspension into 4 liters of deionized waterfollowed by vacuum filtration through Whatman 541 filter paper. Theprocedure of suspension into deionized water followed by vacuumfiltration was repeated several times until the conductivity of thesuspended polymer gel was <1 mS/cm. The polymer gel was then suspendedin deionized water (2 liters) and the mixture was acidified withconcentrated HCl to a pH of <2.5. The mixture was then filtered andtransferred into several Pyrex drying trays. The trays were placed intoa convection oven at 70° C. to dry (24-48 hours). The dried solid wasground to a fine powder using a lab mill with stainless steel blades,and was passed through a sieve (50 mesh) to remove large granules. Theground product was then placed in a vacuum oven at 60° C. and 28 mmHgfor at least 16 hours. Yield=49.9 g TABLE 3 Acetylation reactionsaccording to the procedure Of Example 77 Example Polyamine mol %Acetylation 78 Poly(allylamine) HCl 50 79 Poly(diallylamine) HCl 25 80Poly(diallylamine) HCl 50 81 Poly(diallylamine) HCl 100 

EXAMPLE 82 Alkylation of a Soluble Polymer Preparation of 10 Mol %Dodecyl-Poly(Allylamine)HCl

[0134] Poly(allylamine) Hydrochloride (200 g of 50% aqueous solution,1.07 mol monomer equivalents) was dissolved a mixture of ethanol (213mL) and water (125 mL), and was heated to 70° C. in a 1-liter,round-bottomed flask equipped with an overhead mechanical stirrer, acondenser, and a thermocouple probe. The pH of the solution was broughtto 10.0-10.2 by the addition of NaOH (50% solution). 1-Bromododecane(26.66 g, 0.107 mol) was then added to the stirred solution in oneportion. This mixture was stirred at 70° C. for 20 hours. The solutionpH was checked periodically during this time, and was maintained at10.0-10.2 by the addition of small quantities of 50% NaOH. After the 20hour reaction time had elapsed, the mixture was cooled to roomtemperature and the pH was adjusted to 11.5-12.0 by the addition of a50% NaOH solution. The reaction mixture was then poured into a 5-literbeaker containing 2 liters of methanol stirred with an overheadmechanical stirrer. A fine precipitate was observed as this mixture wasstirred for 30 minutes. The mixture was vacuum filtered through Whatman541 filter paper, and the clear filtrate was acidified with concentratedHCl (pH<1) producing a thick polymer precipitate and a cloudy solution.The cloudy methanol solution was decanted away from the crude solidproduct. The precipitate was dissolved in a minimum amount of water(approx. 300 mL) and acidified with concentrated HCl to a pH of <2. Theaqueous polymer solution was then poured with overhead mechanicalstirring into a 3-liter beaker containing at least 5 volumes (approx.1.5 liters) of methanol (isopropanol can be used in place of methanol inthis step). The polymeric product precipitated as a white solid. Afterstirring for 15 minutes, the precipitate was separated from solution bydecantation and suspended in 2 liters of isopropyl alcohol. The solidwas broken up using a metal spatula and the mixture was stirred for 2hours. The isopropyl alcohol was then removed by decanting and theproduct was again suspended in 2 liters of fresh isopropyl alcohol.After 2 hours of stirring, the solvent was decanted away and the solidproduct was placed in a convection oven at 70° C. to dry (24-48 hours).The dried solid was ground to a fine powder using a lab mill withstainless steel blades, and was passed through a sieve (50 mesh) toremove large granules. The ground product was then placed in a vacuumoven at 60° C. and 28 mmHg for at least 16 hours. Yield=86%.

EXAMPLE 83 Preparation of 0.75 Mol % Crosslinked, 10 Mol %Hexyl-Poly(Allylamine)HCl

[0135] The procedure of Example 82 was used. The 50% aqueous solution ofpoly(allylamine)HCl was replaced with an equivalent amount of a 50%aqueous solution of the polymer product of Example 14. In place of the1-bromododecane, 1-bromohexane (17.66 g, 0.107 mol) was used.

EXAMPLE 84 Preparation of 0.75 Mol % Crosslinked, 10 Mol %Dodecyl-Poly(Allylamine)HCl

[0136] The procedure of Example 82 was used. The 50% aqueous solution ofpoly(allylamine)HCl was replaced with an equivalent amount of a 50%aqueous solution of the polymer product of Example 14.

EXAMPLE 85 Preparation of 0.75 Mol % Crosslinked, 2 Mol %Octadecyl-Poly(Allylamine)HCl

[0137] The procedure of Example 82 was used. The 50% aqueous solution ofpoly(allylamine)HCl was replaced with an equivalent amount of a 50%aqueous solution of the polymer product of Example 14. In place of the1-bromododecane, 1-bromooctadecane (7.13 g, 0.021 mol) was used.

EXAMPLE 86 Preparation of 0.75 Mol % Crosslinked, 25 Mol %Dodecyl-Poly(Allylamine)HCl

[0138] The procedure of Example 82 was used. The amount of dodecylbromide used was 67.3 g, 0.27 mol. The 50% aqueous solution ofpoly(allylamine)HCl was replaced with an equivalent amount of a 50%aqueous solution of the polymer product of Example 14.

EXAMPLE 87 Preparation of 25 Mol % Hexyl-Poly(Allylamine)HCl 0.75 Mol %Epichlorohydrin Crosslinked

[0139] Poly(allylamine) Hydrochloride 0.75% epichlorohydrin crosslinkedfrom Example 14 (200 g of 50% aqueous solution, 1.07 mol monomerequivalents) was dissolved in a mixture of ethanol (213 mL) and water(125 mL), and was heated to 70° C. in a 1-liter, round-bottomed flaskequipped with an overhead mechanical stirrer, a condenser, and athermocouple probe. The pH of the solution was brought to 10.0-10.2 bythe addition of NaOH (50% solution). 1-Bromohexane (44.2 g, 0.27 mol)was then added to the stirred solution in one portion. This mixture wasstirred at 70° C. for 20 hours. The solution pH was checked periodicallyduring this time, and was maintained at 10.0-10.2 by the addition ofsmall quantities of 50% NaOH. After the 20 hour reaction time hadelapsed, the mixture was cooled to room temperature and the pH wasadjusted to 11.5-12.0 by the addition of a 50% NaOH solution. Thereaction mixture was then poured into a 5-liter beaker containing 2liters of methanol stirred with an overhead mechanical stirrer. In thiscase, no precipitate was formed. The methanol solution was evaporated todryness giving a stick solid. The solid was dissolved in 800 mL ofmethanol and 3 liters of hexane was added to precipitate the polymer.After collection by filtration, the polymeric solid was washed with anadditional 1 liter of hexane and collected by filtration. The solidproduct was placed in a convection oven at 70° C. to dry (24-48 hours).The dried solid was ground to a fine powder using a lab mill withstainless steel blades, and was passed through a sieve (50 mesh) toremove large granules. The ground product was then placed in a vacuumoven at 60° C. and 28 mmHg for at least 16 hours. Yield=89.3 g

EXAMPLE 88 Preparation of 50 Mol % Hexyl-Poly(Allylamine)HCl 0.75 Mol %Epichlorohydrin Crosslinked

[0140] Poly(allylamine) Hydrochloride 0.75% epichlorohydrin crosslinkedfrom Example 14 (200 g of 50% aqueous solution, 1.07 mol monomerequivalents) was dissolved in a mixture of ethanol (213 mL) and water(125 mL), and was heated to 70° C. in a 1-liter, round-bottomed flaskequipped with an overhead mechanical stirrer, a condenser, and athermocouple probe. The pH of the solution was brought to 10.0-10.2 bythe addition of NaOH (50% solution). 1-Bromohexane (88.4 g, 0.51 mol)was then added to the stirred solution in one portion. This mixture wasstirred at 70° C. for 20 hours. The solution pH was checked periodicallyduring this time, and was maintained at 10.0-10.2 by the addition ofsmall quantities of 50% NaOH. After the 20 hour reaction time hadelapsed, the mixture was cooled to room temperature giving a whitesuspension of polymer. The white solid was allowed to settle, and thesolution was decanted away. Deionized water (1.5 liters) was added andthe slurry was stirred for 15 minutes. The solid was allowed to settleand the solution was removed by decantation. The solid was thendissolved in 800 mL of isopropanol. Concentrated HCl (120 mL) was addedbut no precipitate was seen. Hexane (3 liters) was added and a whitesolid precipitated from solution. After collection by filtration, thepolymeric solid was washed with an additional 1.5 liters of hexane andcollected by filtration. The solid product was placed in a convectionoven at 70° C. to dry (24-48 hours). The dried solid was ground to afine powder using a lab mill with stainless steel blades, and was passedthrough a sieve (50 mesh) to remove large granules. The ground productwas then placed in a vacuum oven at 60° C. and 28 mmHg for at least 16hours. Yield=117 g

EXAMPLE 89 Preparation of 5 Mol % Dodecyl-Poly(Allylamine)HCl 0.75 Mol %Epichlorohydrin Crosslinked

[0141] Poly(allylamine) Hydrochloride 0.75% epichlorohydrin crosslinkedfrom Example 14 (200 g of 50% aqueous solution, 1.07 mol monomerequivalents) was dissolved in a mixture of ethanol (213 mL) and water(125 mL), and was heated to 70° C. in a 1-liter, round-bottomed flaskequipped with an overhead mechanical stirrer, a condenser, and athermocouple probe. The pH of the solution was brought to 10.0-10.2 bythe addition of NaOH (50% solution). 1-Bromododecane (13.33 g, 0.054mol) was then added to the stirred solution in one portion. This mixturewas stirred at 70° C. for 20 hours. The solution pH was checkedperiodically during this time, and was maintained at 10.0-10.2 by theaddition of small quantities of 50% NaOH. After the 20 hour reactiontime had elapsed, the mixture was cooled to room temperature and the pHwas adjusted to 11.5-12.0 by the addition of a 50% NaOH solution. Thereaction mixture was then poured into a 5-liter beaker containing 2liters of methanol stirred with an overhead mechanical stirrer. A fineprecipitate was observed as this mixture was stirred for 30 minutes. Themixture was vacuum filtered through Whatman 541 filter paper, and theclear filtrate was acidified with concentrated HCl (pH<1) producing athick polymer precipitate and a cloudy solution. The cloudy methanolsolution was decanted away from the crude solid product. Isopropanol(2.5 liters) was added and the solid was broken into small pieces with aspatula. The solid was collected by decantation and washed a second timewith fresh isopropanol. The solid product was placed in a convectionoven at 70° C. to dry (24-48 hours). The dried solid was ground to afine powder using a lab mill with stainless steel blades, and was passedthrough a sieve (50 mesh) to remove large granules. The ground productwas then placed in a vacuum oven at 60° C. and 28 mmHg for at least 16hours. Yield=81 g

EXAMPLE 90 Preparation of 5 Mol % Octadecyl-Poly(Allylamine)HCl 0.75 Mol% Epichlorohydrin Crosslinked

[0142] Poly(allylamine) Hydrochloride 0.75% epichlorohydrin crosslinkedfrom Example 14 (200 g of 50% aqueous solution, 1.07 mol monomerequivalents) was dissolved a mixture of ethanol (213 mL) and water (125mL), and was heated to 70° C. in a 1-liter, round-bottomed flaskequipped with an overhead mechanical stirrer, a condenser, and athermocouple probe. The pH of the solution was brought to 10.0-10.2 bythe addition of NaOH (50% solution). I-Bromooctadecane (17.84 g, 0.054mol) was then added to the stirred solution in one portion. This mixturewas stirred at 70° C. for 20 hours. The solution pH was checkedperiodically during this time, and was maintained at 10.0-10.2 by theaddition of small quantities of 50% NaOH. After the 20 hour reactiontime had elapsed, the mixture was cooled to room temperature and pouredinto a 5-liter beaker containing 2 liters of methanol stirred with anoverhead mechanical stirrer. The mixture was acidified with concentratedHCl (pH<1) producing a thick polymer precipitate. The methanol solutionwas decanted away from the crude solid product, and water (approx. 300mL) was added to disperse the polymeric product into a slurry. Methanol(900 mL) was added giving a dense precipitate. The solution was decantedaway and the polymer was slurried in water a second time andprecipitated with methanol. The solid polymer was then washed with 2.5liters of isopropanol, and then with 1 liter of diethyl ether. The solidproduct was then placed in a convection oven at 70° C. to dry (24-48hours). The dried solid was ground to a fine powder using a lab millwith stainless steel blades, and was passed through a sieve (50 mesh) toremove large granules. The ground product was then placed in a vacuumoven at 60° C. and 28 mmHg for at least 16 hours. Yield=111 g

EXAMPLE 91 Preparation of 10 Mol % Octadecyl-Poly(Allylamine)HCl 0.75Mol % Epichlorohydrin Crosslinked

[0143] The procedure of Example 90 was used. The amount of octadecylbromide used was 35.67 g, 0.107 mol. Yield=124 g

EXAMPLE 92 Preparation of 5 Mol % Docosyl-Poly(Allylamine)HCl 0.75 Mol %Epichlorohydrin Crosslinked

[0144] The procedure of Example 90 was used. The amount of docosylbromide used was 21.03 g, 0.054 mol. Yield=96 g

EXAMPLE 93 Preparation of 10 Mol % Docosyl-Poly(Allylamine)HCl 0.75 Mol% Epichlorohydrin Crosslinked

[0145] The procedure of Example 90 was used. The amount of docosylbromide used was 41.68 g, 0.107 mol. Yield=101 g

EXAMPLE 94 Preparation of 25 Mol % Hexyl-Polyethylenimine HCl

[0146] Polyethylenimine (200 g of a 50% aqueous solution from AldrichChemical Co., 2.32 mol monomer equivalents) was dissolved a mixture ofethanol (213 mL) and water (125 mL), and was heated to 70° C. in a1-liter, round-bottomed flask equipped with an overhead mechanicalstirrer, a condenser, and a thermocouple probe. 1-Bromohexane (95.7 g,0.58 mol) was then added to the stirred solution in one portion. Thismixture was stirred at 70° C. for 20 hours. The solution pH was checkedperiodically during this time, and was maintained at 10.0-10.2 by theaddition of small quantities of 50% NaOH. After the 20 hour reactiontime had elapsed, the mixture was cooled to room temperature and pouredinto a 20-liter bucket containing 3 liters of methanol stirred with anoverhead mechanical stirrer. The mixture was acidified with concentratedHCl (pH<2), but no precipitate was formed. Isopropanol (6 liters) wasadded, giving a small amount of precipitate. Diethyl ether was thenadded (3 liters) and the crude product precipitated. The solvent wasdecanted away from the product. The crude product was then redispersedin 750 mL of deionized water. The pH was adjusted to <2 usingconcentrated HCl. Acetonitrile (5 liters) was then added to precipitatethe polymer. The solid was collected by decantation and washed with 2liters of isopropanol. The solid product was then placed in a convectionoven at 70° C. to dry (24-48 hours). The dried solid was ground to afine powder using a lab mill with stainless steel blades, and was passedthrough a sieve (50 mesh) to remove large granules. The ground productwas then placed in a vacuum oven at 60° C. and 28 mmHg for at least 16hours. Yield=172 g

EXAMPLE 95 Preparation of 50 Mol % Hexyl-Polyethylenimine HCl

[0147] Polyethylenimine (200 g of a 50% aqueous solution from AldrichChemical Co., 2.32 mol monomer equivalents) was dissolved in a mixtureof ethanol (213 mL) and water (125 mL), and was heated to 70° C. in a1-liter, round-bottomed flask equipped with an overhead mechanicalstirrer, a condenser, and a thermocouple probe. 1-Bromohexane (191.5 g,1.16 mol) was then added to the stirred solution in one portion. Thismixture was stirred at 70° C. for 20 hours. The solution pH was checkedperiodically during this time, and was maintained at 10.0-10.2 by theaddition of small quantities of 50% NaOH. After the 20-hour reactiontime had elapsed, the mixture was cooled to room temperature and pouredinto a 20-liter bucket containing 2 liters of methanol stirred with anoverhead mechanical stirrer. The mixture was acidified with concentratedHCl (pH<2), but no precipitate was formed. Diethyl ether was then added(3 liters) and the crude product precipitated. The solvent was decantedaway from the product. The crude product was then redispersed in ethanol(3 liters). The pH was adjusted to >11.5 using concentrated NaOH. Thefree base polymer dissolved, leaving a suspension of salts. The mixturewas vacuum filtered through Whatman 541 filter paper, and the clearfiltrate was acidified with concentrated HCl (pH<1). Diethyl ether wasthen added to precipitate the product, which was then collected bydecantation. The solid product was then placed in a convection oven at70° C. to dry (24-48 hours). The dried solid was ground to a fine powderusing a lab mill with stainless steel blades, and was passed through asieve (50 mesh) to remove large granules. The ground product was thenplaced in a vacuum oven at 60° C. and 28 mmHg for at least 16 hours.Yield=110 g

EXAMPLE 96 Preparation of 5 Mol % Dodecyl-Polyethylenimine HCl

[0148] Polyethylenimine (200 g of a 50% aqueous solution from AldrichChemical Co., 2.32 mol monomer equivalents) was dissolved in a mixtureof ethanol (213 mL) and water (125 mL), and was heated to 70° C. in a1-liter, round-bottomed flask equipped with an overhead mechanicalstirrer, a condenser, and a thermocouple probe. 1-Bromododecane (28.9 g,0.116 mol) was then added to the stirred solution in one portion. Thismixture was stirred at 70° C. for 20 hours. The solution pH was checkedperiodically during this time, and was maintained at 10.0-10.2 by theaddition of small quantities of 50% NaOH. After the 20 hour reactiontime had elapsed, the mixture was cooled to room temperature and pouredinto a 20-liter bucket containing 2 liters of methanol stirred with anoverhead mechanical stirrer. The mixture was acidified with concentratedHCl (pH<2), resulting in the precipitation of some polymer. Isopropanolwas then added (3 liters) and the crude product precipitated. Thesolvent was decanted away from the product. The crude product was thenredispersed in water (750 mL) and methanol (400 mL) and the pH wasadjusted to <2 using concentrated HCl. Isopropanol (6 liters) was thenadded to precipitate the product, which was collected by decantation.The solid product was then placed in a convection oven at 70° C. to dry(24-48 hours). The dried solid was ground to a fine powder using a labmill with stainless steel blades, and was passed through a sieve (50mesh) to remove large granules. The ground product was then placed in avacuum oven at 60° C. and 28 mmHg for at least 16 hours. Yield=146 g

EXAMPLE 97 Preparation of 10 Mol % Dodecyl-Polyethylenimine HCl

[0149] Polyethylenimine (200 g of a 50% aqueous solution from AldrichChemical Co., 2.32 mol monomer equivalents) was dissolved in a mixtureof ethanol (213 mL) and water (125 mL), and was heated to 70° C. in a1-liter, round-bottomed flask equipped with an overhead mechanicalstirrer, a condenser, and a thermocouple probe. 1-Bromododecane (57.8 g,0.232 mol) was then added to the stirred solution in one portion. Thismixture was stirred at 70° C. for 20 hours. The solution pH was checkedperiodically during this time, and was maintained at 10.0-10.2 by theaddition of small quantities of 50% NaOH. After the 20 hour reactiontime had elapsed, the mixture was cooled to room temperature and pouredinto a 20-liter bucket containing 2 liters of methanol stirred with anoverhead mechanical stirrer. The mixture was acidified with concentratedHCl (pH<2), resulting in the precipitation of some polymer. Isopropanolwas then added (3 liters) and the crude product precipitated. Thesolvent was decanted away from the product. The crude product was thenredispersed in water (750 mL) and methanol (400 mL) and the pH wasadjusted to <2 using concentrated HCl. Isopropanol (6 liters) was thenadded to precipitate the product, which was collected by decantation.The solid product was then placed in a convection oven at 70° C. to dry(24-48 hours). The dried solid was ground to a fine powder using a labmill with stainless steel blades, and was passed through a sieve (50mesh) to remove large granules. The ground product was then placed in avacuum oven at 60° C. and 28 mmHg for at least 16 hours. Yield=119 g

EXAMPLE 98 Preparation of 25 Mol % Dodecyl-Polyethylenimine HCl

[0150] Polyethylenimine (200 g of a 50% aqueous solution from AldrichChemical Co., 2.32 mol monomer equivalents) was dissolved in a mixtureof ethanol (213 mL) and water (125 mL), and was heated to 70° C. in a1-liter, round-bottomed flask equipped with an overhead mechanicalstirrer, a condenser, and a thermocouple probe. 1-Bromododecane (144.6g, 0.58 mol) was then added to the stirred solution in one portion. Thismixture was stirred at 70° C. for 20 hours. The solution pH was checkedperiodically during this time, and was maintained at 10.0-10.2 by theaddition of small quantities of 50% NaOH. After the 20 hour reactiontime had elapsed, the mixture was cooled to room temperature and pouredinto a 20-liter pail containing 5 liters of ethanol stirred with anoverhead mechanical stirrer. The mixture was acidified with concentratedHCl (pH<2), resulting in the precipitation of the crude product. Thesolvent was decanted, and the crude product was then redissolved inwater (750 mL). The pH was adjusted to <2 using concentrated HCl.Isopropanol (6 liters) was then added to precipitate the product, whichwas collected by decantation. The solid product was then placed in aconvection oven at 70° C. to dry (24-48 hours). The dried solid wasground to a fine powder using a lab mill with stainless steel blades,and was passed through a sieve (50 mesh) to remove large granules. Theground product was then placed in a vacuum oven at 60° C. and 28 mmHgfor at least 16 hours. Yield=197 g

EXAMPLE 99 Preparation of 2.5 Mol % Octadecyl-Polyethylenimine HCl

[0151] Polyethylenimine (200 g of a 50% aqueous solution from AldrichChemical Co., 2.32 mol monomer equivalents) was dissolved in a mixtureof ethanol (213 mL) and water (125 mL), and was heated to 70° C. in a1-liter, round-bottomed flask equipped with an overhead mechanicalstirrer, a condenser, and a thermocouple probe. 1-bromooctadecane (19.3g, 0.058 mol) was then added to the stirred solution in one portion.This mixture was stirred at 70° C. for 20 hours. The solution pH waschecked periodically during this time, and was maintained at 10.0-10.2by the addition of small quantities of 50% NaOH. After the 20 hourreaction time had elapsed, the mixture was cooled to room temperatureand poured into a 20-liter bucket containing 3.5 liters of methanolstirred with an overhead mechanical stirrer. The mixture was acidifiedwith concentrated HCl (pH<2), resulting in the precipitation of thecrude product. The solvent was decanted, and the crude product was thenredissolved in water (1100 mL). The pH was adjusted to <2 usingconcentrated HCl. Isopropanol (6 liters) was then added to precipitatethe product, which was collected by decantation. The solid was washedwith another 2 liters of clean isopropanol and collected by decantation.The solid product was then placed in a convection oven at 70° C. to dry(24-48 hours). The dried solid was ground to a fine powder using a labmill with stainless steel blades, and was passed through a sieve (50mesh) to remove large granules. The ground product was then placed in avacuum oven at 60° C. and 28 mmHg for at least 16 hours. Yield=169 g

EXAMPLE 100 Preparation of 5 Mol % Octadecyl-Polyethylenimine HCl

[0152] The procedure of Example 99 was used. The amount of1-bromooctadecane used 38.7 g, 0.116 mol. Yield=198 g

EXAMPLE 101 Preparation of 2 Mol % Docosyl-Polyethylenimine HCl

[0153] Polyethylenimine (200 g of a 50% aqueous solution from AldrichChemical Co., 2.32 mol monomer equivalents) was dissolved in a mixtureof ethanol (213 mL) and water (125 mL), and was heated to 70° C. in a1-liter, round-bottomed flask equipped with an overhead mechanicalstirrer, a condenser, and a thermocouple probe. 1-Bromodocosane (18.1 g,0.046 mol) was then added to the stirred solution in one portion. Thismixture was stirred at 70° C. for 20 hours. The solution pH was checkedperiodically during this time, and was maintained at 10.0-10.2 by theaddition of small quantities of 50% NaOH. After the 20 hour reactiontime had elapsed, the mixture was cooled to room temperature and pouredinto a 20-liter bucket containing 2.4 liters of ethanol stirred with anoverhead mechanical stirrer. The mixture was acidified with concentratedHCl (pH<2), resulting in the precipitation the crude product. Thesolvent was decanted, and the crude product was then redissolved inwater (700 mL). The pH was adjusted to <2 using concentrated HCl.Isopropanol (6 liters) was then added to precipitate the product, whichwas collected by decantation. The solid was washed with another 2 litersof clean isopropanol and collected by decantation. The solid product wasthen placed in a convection oven at 70° C. to dry (24-48 hours). Thedried solid was ground to a fine powder using a lab mill with stainlesssteel blades, and was passed through a sieve (50 mesh) to remove largegranules. The ground product was then placed in a vacuum oven at 60° C.and 28 mmHg for at least 16 hours. Yield=154 g

EXAMPLE 102 Preparation of 5 Mol % Docosyl-Polyethylenimine HCl

[0154] The procedure of Example 101 was used. The amount of1-bromodocosane used was 45.2 g, 0.116 mol. Yield=160 g

EXAMPLE 103 Preparation of 5 Mol % dodecyl-poly(diallylamine)HCl 0.75Mol % Epichlorohydrin Crosslinked

[0155] Poly(diallylamine) Hydrochloride 0.75% epichlorohydrincrosslinked from Example 15 (60 g, 0.45 mol monomer equivalents) wasdispersed in ethanol (500 mL) in a 1-liter, round-bottomed flaskequipped with an overhead mechanical stirrer, a condenser, and athermocouple probe. NaOH was added (30 g of a 50% solution), along withdeionized water (200 mL) and the mixture was heated to 70° C. The pH ofthe solution was brought to 10.0-10.2 by the addition of NaOH (50%solution). 1-Bromododecane (5.6 g, 0.023 mol) was then added to thestirred solution in one portion. This mixture was stirred at 70° C. for20 hours. The solution pH was checked periodically during this time, andwas maintained at 10.0-10.2 by the addition of small quantities of 50%NaOH. After the 20 hour reaction time had elapsed, the mixture wascooled to room temperature and poured into a 5-liter beaker containing 3liters of deionized water stirred with an overhead mechanical stirrer.The crude polymeric product precipitated from solution and was collectedby decantation. The crude product was added to a mixture of 300 mLdeionized water and 300 mL ethanol. The mixture was acidified withconcentrated HCl (pH<1) and stirred for at least 2 hours. Isopropanol (3liters) was then added to precipitate the product. The solid polymer wasthen washed with clean isopropanol (2 liters). The solid product wasthen placed in a convection oven at 70° C. to dry (24-48 hours). Thedried solid was ground to a fine powder using a lab mill with stainlesssteel blades, and was passed through a sieve (50 mesh) to remove largegranules. The ground product was then placed in a vacuum oven at 60° C.and 28 mmHg for at least 16 hours. Yield=27 g

EXAMPLE 104 Preparation of 10 Mol % Dodecyl-Poly(Diallylamine)HCl 0.75Mol % Epichlorohydrin Crosslinked

[0156] The procedure of Example 104 was used. The amount of1-bromododecane used was 11.2 g, 0.045 mol. Yield=46 g

EXAMPLE 105 Preparation of 25 Mol % Dodecyl-Poly(Diallylamine)HCl 0.75Mol % Epichlorohydrin Crosslinked

[0157] The procedure of Example 104 was used. The amount of1-bromododecane used was 28.2 g, 0.113 mol. Yield=52 g

EXAMPLE 106 Preparation of 5 Mol % Hexyl-Poly(Diallylamine)HCl 0.75 Mol% Epichlorohydrin Crosslinked

[0158] The procedure of Example 104 was used. The amount of1-bromohexane used was 3.72 g, 0.023 mol.

EXAMPLE 107 Preparation of 10 Mol % Hexyl-Poly(Diallylamine)HCl 0.75 Mol% Epichlorohydrin Crosslinked

[0159] The procedure of Example 104 was used. The amount of1-bromohexane used was 7.43 g, 0.045 mol. Yield=36 g

EXAMPLE 108 Preparation of 25 Mol % Hexyl-Poly(Diallylamine)HCl 0.75 Mol% Epichlorohydrin Crosslinked

[0160] The procedure of Example 104 was used. The amount of1-bromohexane used was 18.65 g, 0.113 mol. Yield=49 g

EXAMPLE 109 Preparation of 50 Mol % Hexyl-Poly(Diallylamine)HCl 0.75 Mol% Epichlorohydrin Crosslinked

[0161] The procedure of Example 104 was used. The amount of1-bromohexane used was 38.0 g, 0.230 mol. Yield=67 g

EXAMPLE 110 Preparation of 2 Mol % Octadecyl-Poly(Diallylamine)HCl 0.75Mol % Epichlorohydrin Crosslinked

[0162] The procedure of Example 104 was used. The amount of1-bromooctadecane used was 3.0 g, 0.009 mol. Yield=23 g

EXAMPLE 111 Preparation of 5 Mol % Octadecyl-Poly(Diallylamine)HCl 0.75Mol % Epichlorohydrin Crosslinked

[0163] The procedure of Example 104 was used. The amount of1-bromooctadecane used was 7.7 g, 0.023 mol. Yield=35 g

EXAMPLE 112 Preparation of 10 Mol % Octadecyl-Poly(Diallylamine)HCl 0.75Mol % Epichlorohydrin Crosslinked

[0164] The procedure of Example 104 was used. The amount of1-bromooctadecane used was 15.0 g, 0.045 mol. Yield=35 g

EXAMPLE 113 Preparation of Copolymer of Acrylamide (20 Mole%)/Trimethylaminoethyl Acrylchloride Q Salt (TMAEAC) (78 Mole%)/Octadecylacrylate (2 Mole %)

[0165] To a 1-liter, three-necked flask equipped with condenser and stirbar, were added trimethylaminoethyl acrylchloride quaternary salt(TMAEAC) 50% aqueous solution (150.90 g of 50% solution, 390 mmoles, 78mole %), acrylamide (7.11 g, 100 mmoles, 20 mole %), octadecyl acrylate(3.25 g, 10 mmoles, 2 mole %) and isopropanol (400 mL). The mixture waspurged with nitrogen for 10 min before the addition of a radicalinitiator, AIBN (330 mg, 2 mmoles). The mixture was heated to 70° C. for16 hours. At the end of the 16 hour reaction time, the reaction mixturewas allowed to cool to room temperature and poured into a beakercontaining isopropanol (1 liter). The polymer was precipitated out as awhite solid, which was collected and ground to small pieces in a blenderusing isopropanol as a solvent. The pieces were collected by filtrationand the polymer was dried under vacuum at 60° C. for 2 days. Thematerial was ground to a fine powder (82 g), which was used for the invitro and in vivo studies.

[0166] The following polymers of Table 4 with varying composition ofacrylamide, TMAEAC and octadecyl acrylate were prepared using the aboveprocedure. TABLE 4 Acrylamide TMAEAC Octadecyl Example No. (mole %)(mole %) acrylate (mole %) 114 20 75 5 115 20 70 10  116 35 63 2 117 3560 5 118 35 55 10  119 50 48 2 120 50 45 5 121 50 40 10  122  0 98 2 123 0 95 5 124  0 90 10 

EXAMPLE 125 Preparation of Copolymer of Acrylamide (20 Mole%)/Trimethylaminoethyl Acrylchloride Q Salt (TMAEAC) (75 Mole%)/Dodecylacrylate (5 Mole %)

[0167] To a 1-liter, three-necked flask equipped with condenser and stirbar, were added trimethylaminoethyl acrylchloride quaternary salt(TMAEAC) 50% aqueous solution (145.80 g of 50% solution, 375 mmoles, 75mole %), acrylamide (7.11 g, 100 mmoles, 20 mole %), dodecyl acrylate(6.01 g, 25 mmoles, 5 mole %) and isopropanol (400 mL). The mixture waspurged with nitrogen for 10 min before the addition of a radicalinitiator, AIBN (330 mg, 2 mmoles). The mixture was heated to 70° C. for16 hours. The reaction mixture was allowed to cool to room temperatureand poured into a beaker containing isopropanol (1 liter). The polymerwas precipitated out as a white solid, which was collected and ground tosmall pieces in a blender using isopropanol as a solvent. The pieceswere collected by filtration and the polymer was dried under vacuum at60° C. for 2 days. The material was ground to a fine powder (80 g),which was used for the in vitro and in vivo studies. The polymers ofTable 5 were prepared using the above procedure. TABLE 5 ExampleAcrylamide TMAEAC Dodecylacrylate No. (mole %) (mole %) (mole %) 126 2070 10 127 20 55 25 128 35 60  5 129 35 55 10 130 35 40 25 131 50 45  5132 50 40 10 133 50 25 25 134  0 95  5 135  0 90 10 136  0 75 25 137  098  2 138 20 78  2 139 35 63  2 140 50 48  2

EXAMPLE 141 Preparation of Copolymer of Acrylamide (20 Mole%)/Trimethylaminoethyl Acrylchloride Q Salt (TMAEAC) (80 Mole %)

[0168] To a 1-liter, three-necked flask equipped with condenser and stirbar, were added trimethylaminoethyl acrylchloride quaternary salt(TMAEAC) 50% aqueous solution (154.76 g of 50% solution, 400 mmoles, 80mole %), acrylamide (7.11 g, 100 mmoles, 20 mole %), and isopropanol(400 mL). The mixture was purged with nitrogen for 10 min before theaddition of a radical initiator, AIBN (330 mg, 2 mmoles). The mixturewas heated to 70° C. for 16 hours. At the end of reaction, reactionmixture was allowed to cool to room temperature and poured into a beakercontaining isopropanol (1 liter). The polymer was precipitated out as awhite solid, which was collected and ground to small pieces in a blenderusing isopropanol as a solvent. The pieces were collected by filtrationand the polymer was dried under vacuum at 60° C. for 2 days. Thematerial was ground to a fine powder (80 g), which was used for the invitro and in vivo studies.

[0169] The polymers of Table 6 were prepared using the above procedureTABLE 6 Example No. Acrylamide (mole %) TMAEAC (mole %) 142  0 100  14310 90 141 20 80 144 35 65 145 50 50

EXAMPLE 146 Preparation of Methylenebisacrylamide (4 Mole %)Cross-Linked Polymers of Acrylamide (20 Mole %)/TrimethylaminoethylAcrylchloride Q Salt (TMAEAC) (78 Mole %)/Octadecyl Acrylate (2 Mole %)

[0170] To a 1-liter, three-necked flask equipped with condenser and stirbar, were added trimethylaminoethyl acrylchloride quaternary salt(TMAEAC) 50% aqueous solution (150.90 g of 50% solution, 390 mmoles, 78mole %), acrylamide (7.11 g, 100 mmoles, 20 mole %), octadecyl acrylate(3.25 g, 10 mmoles, 2 mole %), methylenebisacrylamide (3.08 g, 20mmoles, 4 mole %) and ethanol (300 mL) were added. The mixture waspurged with nitrogen for 10 min before the addition of a radicalinitiator, AIBN (330 mg, 2 mmoles). The mixture was heated to 70° C. for16 hours. At the end of reaction, reaction mixture was allowed to coolto room temperature and poured into a beaker containing isopropanol (1liter). The polymer was precipitated out as a white solid, which wascollected and ground to small pieces in a blender using isopropanol as asolvent. The pieces were collected by filtration and the polymer wasdried under vacuum at 60° C. for 2 days. The material was ground to afine powder (84 g), which was used for the in vitro and in vivo studies.

[0171] The compounds of Table 7 were prepared using the above procedure.TABLE 7 Octadecyl Example Acrylamide TMAEAC acrylate Methylenebisacryl-No. (mole %) (mole %) (mole %) amide (mole %) 147 20 75 5 4 148 20 7010  4 149 35 63 2 4 150 35 60 5 4 151 35 55 10  4 152 50 48 2 4 153 5045 5 4 154 50 40 10  4 155  0 98 2 4 156  0 95 5 4 157  0 90 10  4

EXAMPLE 158 Preparation of Methylenebisacrylamide (4 Mole %)Cross-Linked Polymer of Acrylamide (20 Mole %)/TrimethylaminoethylAcrylchloride Q Salt (TMAEAC) (75 Mole %)/Dodecylacrylate (5 Mole %)

[0172] To a 1-liter, three-necked flask equipped with condenser and stirbar, were added trimethylaminoethyl acrylchloride quaternary salt(TMAEAC) 50% aqueous solution (145.80 g of 50% solution, 375 mmoles, 75mole %), acrylamide (7.11 g, 100 mmoles, 20 mole %), dodecylacrylate(6.01 g, 25 mmoles, 5 mole %), methylenebisacrylamide (3.08 g, 20mmoles, 4 mole %) and ethanol (300 mL). The mixture was purged withnitrogen for 10 min before the addition of a radical initiator, AIBN(330 mg, 2 mmoles). The mixture was heated to 70° C. for 16 hours. Thereaction mixture was allowed to cool to room temperature and poured intoa beaker containing isopropanol (1 liter). The polymer was precipitatedout as a white solid, which was collected and ground to small pieces ina blender using isopropanol as a solvent. The pieces were collected byfiltration and the polymer was dried under vacuum at 60° C. for 2 days.The material was ground to a fine powder (80 g), which was used for thein vitro and in vivo studies. The following cross-linked polymers wereprepared.

[0173] The polymers of Table 8 were prepared using the above procedure.TABLE 8 Dodecyl- Example Acrylamide TMAEAC acrylate Methylenebisacryl-No. (mole %) (mole %) (mole %) amide (mole %) 159 20 70 10 4 160 20 5525 4 161 35 60  5 4 162 35 55 10 4 163 35 40 25 4 164 50 45  5 4 165 5040 10 4 166 50 25 25 4 167  0 95  5 4 168  0 90 10 4 169  0 75 25 4

EXAMPLE 170 Preparation of Copolymer of Methacrylamide (20 Mol%)/Trimethylaminoethyl Methachloride Quaternary Salt (TMAEMC) (78 Mol%)/Octadecyl Methacrylate (2 Mol %)

[0174] To a 1 liter, three necked, round-bottomed flask equipped withcondenser, stir bar, heating mantle (with J-Kem temperature controller)and nitrogen bubbler was added: trimethylammonioethyl methacrylatechloride (TMAEMC) ˜75% aqueous 87.19 g (420.39 mmol), octadecylmethacrylate (3.65 g, 10.80 mmol), methacrylamide (9.16 g, 107.76 mmol),and ethanol (300 mL). The total amount of monomer solids should be 100g. Nitrogen was allowed to bubble through the room temperature monomermixture for at least 20 minutes before adding 0.275 g AIBN[2,2′-azobis(2-methyl-propionitrile)]. At this point the nitrogen wasset to blanket the mixture and the heat was turned on to 70° C. Thereaction was allowed to heat for 22 hours at 70° C. While the polymerwas still warm it was poured from the flask into a Nalgene bucket andallowed to stand for at least three hours in each of four 1-literwashings of isopropanol. Once the polymer became slightly rigid/rubberyit was broken up into small chunks using a blender (with isopropanol asthe liquid). The granular product was filtered and washed with moreisopropanol and placed in a crystallizing dish in a 70° C. convectionoven for two days. After this time, the product was removed and groundto a fine powder using a grinder and placed back in the oven for twomore days. GPC analysis of octadecyl methacrylate containing polymersshows MW ranges form 100K-150K with polydispersities ranging from 2.5-5.The following table gives general mole percent compositions of polymersprepared in this fashion. The polymers of Table 9 were prepared usingthe above procedure. TABLE 9 Octadecyl Example Methacrylamide TMAEMCMethacrylate No. (mole %) (mole %) (mole %) 171  0 98 2 172  0 95 5 173 0 90 10  174  0 85 15  175  0 80 20  170 20 78 2 176 20 75 5 177 20 7010  178 35 63 2 179 35 60 5 180 35 55 10  181 50 48 2 182 50 45 5 183 5040 10 

EXAMPLE 184 Preparation of Copolymer of Methacrylamide (20 Mol%)/Trimethylaminoethyl Methacry-Chloride Quaternary Salt (TMAEMC) (75Mol %) Idodecyl Methacrylate (5 Mol %)

[0175] To a 1-liter, three-necked, round-bottomed flask equipped withcondenser, stir bar, heating mantle (with J-Kem temperature controller)and nitrogen bubbler was added: trimethylammonioethyl methacrylatechloride (TMAEMC) ˜75% aqueous (83.97 g, 404.86 mmol), dodecylmethacrylate (6.85 g, 26.96 mmol), methacrylamide (9.18 g, 108.0 mmol),and ethanol (300 mL). The total amount of monomer solids should be 100g. Nitrogen was allowed to bubble through the room temperature monomermixture for at least 20 minutes before adding 0.275 g AIBN[2,2′-azobis(2-methyl-propionitrile)]. At this point, the nitrogen wasset to blanket the mixture and the heat was turned on to 70° C. Thereaction was allowed to heat for 22 hours at 70° C. While the polymerwas still warm it was poured from the flask into a Nalgene bucket andallowed to stand for at least three hours in each of four 1-literwashings of isopropanol. Once the polymer became slightly rigid/rubberyit was broken up into small chunks using a blender (with isopropanol asthe liquid). The granular product was filtered and washed with moreisopropanol and placed in a crystallizing dish in a 70° C. convectionoven for two days. After this time, the product was removed and groundto a fine powder using a grinder and placed back in the oven for twomore days. GPC analysis of dodecyl methacrylate containing polymersshows MW ranges form 170-190K with polydispersities ranging from2.3-2.8. Table 10 gives general mole percent compositions of polymersprepared in this fashion.

[0176] The polymers of Table 10 were prepared using the above procedure.TABLE 10 Example Methacrylamide TMAEMC Dodecyl metrhacrylate No. (mole%) (mole %) (mole %) 185  0 95  5 186  0 90 10 187  0 75 25 184 20 75  5188 20 70 10 189 20 55 25 190 35 60  5 191 35 55 10 192 35 40 25 193 5045  5 194 50 40 10 195 50 25 25

EXAMPLE 196 Preparation of Copolymer of TMAEMC (80 Mol %)/Methacrylamide(20 Mol %)

[0177] To a 1-liter, three-necked, round-bottomed flask equipped withcondenser, stir bar, heating mantle (with J-Kem temperature controller)and nitrogen bubbler was added: trimethylammonioethyl methacrylatechloride (TMAEMC) ˜75% aqueous, (90.71 g, 437.36 mmol), methacrylamide(9.29 g, 109.29 mmol), and ethanol (300 mL). The total amount of monomersolids should be 100 g. Nitrogen was allowed to bubble through the roomtemperature monomer mixture for at least 20 minutes before adding 0.275g AIBN [2,2′-azobis(2-methyl-propionitrile)]. At this point, thenitrogen was set to blanket the mixture and the heat was turned on to70° C. The reaction was allowed to heat for 22 hours at 70° C. While thepolymer was still warm it was poured from the flask into a Nalgenebucket and allowed to stand for at least three hours in each of four1-liter washings of isopropanol. Once the polymer became slightlyrigid/rubbery it was broken up into small chunks using a blender (withisopropanol as the liquid). The granular product was filtered and washedwith more isopropanol and placed in a crystallizing dish in a 70° C.convection oven for two days. After this time the product was removedand ground to a fine powder using a grinder and placed back in the ovenfor two more days. Table 11 gives general mole percent compositions ofpolymers prepared in this fashion.

[0178] The polymers of Table 11 were prepared using the above procedure.TABLE 11 Example No. Methacrylamide (mole %) TMAEMC (mole %) 197  0 100 196 20 80 198 35 65 199 50 50

EXAMPLE 200 Preparation of Methacrylate Methylenebismethacrylamide (2Mole %) Cross-Linked Polymer of Methacrylamide (20 Mol %)/TMAEMC (78 Mol%)/Octadecyl Methacrylate (2 Mol %)

[0179] To a 1-liter, three-necked, two-part reaction flask equipped withcondenser, mechanical stirrer, water bath, and nitrogen bubbler wasadded trimethylammonioethyl methacrylate chloride (TMAEMC), ˜75% aqueous(87.19 g, 420.39 mmol) octadecyl methacrylate (3.65 g, 10.80 mmol),methacrylamide (9.16 g, 107.76 mmol), and ethanol (400 mL). The totalamount of monomer solids should be 100 g. To this was added anadditional 2 mole percent (of total monomers)N,N′-methylenebismethacrylamide (1.96 g, 10.76 mmol). Nitrogen wasallowed to bubble through the room temperature monomer mixture for atleast 20 minutes before adding 0.275 g AIBN[2,2′-azobis(2-methyl-propionitrile)]. At this point, the nitrogen wasset to blanket the mixture and the heat was turned on to 70° C. Once thepolymer began to gel the stirring was turned off; total heating time at70° C. was approximately 5 hours. The polymer was then allowed cool downto room temperature and stand overnight. The gelled product was scoopedout of the flask and swollen to a clear gel in a 500 mL isopropanol/1000mL water mixture. The gel was washed 6×with 1000 mL isopropanolfiltering over a 50-mesh sieve. Once the polymer became slightlyrigid/rubbery it was broken up into small chunks using a blender (withisopropanol as the liquid). The product was filtered over a sieve, wrungout, and placed in a drying dish in a 70° C. convection oven for twodays. After this time, the product was removed and ground to a finepowder using a grinder and placed back in the oven for two more days ina glass crystallizing dish. Table 12 gives general mole percentcompositions of polymers prepared in this fashion.

[0180] The polymers of Table 12 were prepared using the above procedure.TABLE 12 Methacryl- Octadecyl Methylenebismeth- Example amide TMAEMCMethacrylate acrylamide No. (mole %) (mole %) (mole %) (mole %) 201  098  2 2 202  0 95  5 2 203  0 90 10 2 204  0 85 15 2 205  0 80 20 2 20020 78  2 2 206 20 75  5 2 207 20 70 10 2 208 35 63  2 2 209 35 60  5 2210 35 55 10 2 211 50 48  2 2 212 50 45  5 2 213 50 40 10 2

EXAMPLE 214 Preparation of Methacrylatemethylene-Bismethacrylamide (2Mole %) Cross-Linked Polymer of Methacrylamide (20 Mol %)/TMAEMC (75 Mol%)/Dodecyl Methacrylate (5 Mol %)

[0181] To a 11-liter, three-necked, two-part reaction flask equippedwith condenser, mechanical stirrer, water bath, and nitrogen bubbler wasadded: trimethylammonioethyl methacrylate chloride (TMAEMC), ˜75%aqueous solution (83.97 g, 404.86 mmol), dodecyl methacrylate (6.85 g,26.96 mmol), methacrylamide (9.18 g, 108.0 mmol) and ethanol (400 mL).The total amount of monomer solids should be 100 g. To this was added anadditional 2 mole percent (of total monomers)N,N′-methylenebismethacrylamide (1.96 g, 10.79 mmol). Nitrogen wasallowed to bubble through the room temperature monomer mixture for atleast 20 minutes before adding 0.275 g AIBN[2,2′-azobis(2-methyl-propionitrile)]. At this point, the nitrogen wasset to blanket the mixture and the heat was turned on to 70° C. Once thepolymer began to gel the stirring was turned off; total heating time at70° C. was approximately 5 hours. The polymer was then allowed cool downto room temperature and stand overnight. The gelled product was scoopedout of the flask and swollen to a clear gel in a 500 mL isopropanol/1000mL water mixture. The gel was washed 6× with 1000 mL of isopropanolfiltering over a 50-mesh sieve. Once the polymer became slightlyrigid/rubbery it was broken up into small chunks using a blender (withisopropanol as the liquid). The product was filtered over a sieve, wrungout, and placed in a drying dish in a 70° C. convection oven for twodays. After this time, the product was removed and ground to a finepowder using a grinder and placed back in the oven for two more days ina glass crystallizing dish. Table 13 gives general mole percentcompositions of polymers prepared in this fashion. TABLE 3 Methacryl-Dodecyl Methylenebismeth- Example amide TMAEC methacrylate acrylamideNo. (mole %) (mole %) (mole %) (mole %) 215  0 95  5 2 216  0 90 10 2217  0 75 25 2 214 20 75  5 2 218 20 70 10 2 219 20 55 25 2 220 35 60  52 221 35 55 10 2 222 35 40 25 2 223 50 45  5 2 224 50 40 10 2 225 50 2525 2

EXAMPLE 226 Preparation of Methacrylatemethylenebismethacrylamide (2Mole %) Cross-Linked Polymer of Methacrylamide (20 Mol %)/TMAEMC (80 Mol%)

[0182] To a 1-liter, three-necked, two-part reaction flask equipped withcondenser, mechanical stirrer, water bath, and nitrogen bubbler wasadded: trimethylammonioethyl methacrylate chloride (TMAEMC) ˜75% aqueoussolution (90.71 g, 437.36 mmol), methacrylamide 9.29 g (109.29 mmol),and ethanol (400 mL). The total amount of monomer solids should be 100g. To this was added an additional 2 mole percent (of total monomers)N,N′-methylenebismethacrylamide (1.989 g, 10.93 mmol). Nitrogen wasallowed to bubble through the room temperature monomer mixture for atleast 20 minutes before adding 0.275 g AIBN[2,2′-azobis(2-methyl-propionitrile)]. At this point, the nitrogen wasset to blanket the mixture and the heat was turned on to 70° C. Once thepolymer began to gel the stirring is turned off; total heating time at70° C. was approximately 5 hours. The polymer was then allowed cool downto room temperature and stand overnight. The gelled product was scoopedout of the flask and swollen to a clear gel in a 500 mL isopropanol/1000mL of water mixture. The gel was washed 6×with 1000 mL isopropanolfiltering over a 50-mesh sieve. Once the polymer became slightlyrigid/rubbery it was broken up into small chunks using a blender (withisopropanol as the liquid). The spongy product is filtered over a sieve,pressed dry, and placed in a drying dish in a 70° C. convection oven fortwo days. After this time the product was removed and ground to a finepowder using a grinder and placed back in the oven for two more days ina glass crystallizing dish. Table 14 gives general mole percentcompositions of polymers prepared in this fashion. TABLE 14 ExampleMethacrylamide TMAEMC Methylenebismeth- No. (mole %) (mole %) Acrylamide(mole %) 227  0 100  2 226 20 80 2 228 35 65 2 229 50 50 2

[0183] Method for Determining Binding of Emulsion Particles byLipid-Binding Polymers Using an Olive Oil Emulsion with PhysiologicalEmulsifiers

[0184] Preparation of Olive Oil Emulsion for Lipid-Binding Assay

[0185] Emulsifier Solution

[0186] Egg yolk lecithin 2.54 mmol (2.00 g) and cholesterol 1.25 mmol(0.483 g) were dissolved in 100 mL of chloroform in a 1-liter,round-bottomed flask and the solvent was removed rapidly using a rotaryevaporator. A coating of lecithin and cholesterol resulted, adhering tothe walls of the flask. This film was held under vacuum for 12 hours.The sodium salts of the following bile acids were then added to theflask: glycocholic 1.217 g (2.496 mmol), taurocholic 0.895 g (1.664mmol), glycodeoxycholic 1.766 g (3.744 mmol), taurodeoxycholic 1.302 g(2.496 mmol). An aqueous buffer consisting of 0.1M2-[N-morpholino]ethanesulfonic acid (MES) and 0.1M sodium chloride wasprepared and the pH was adjusted with 50% NaOH to pH=6.5.1 liter of thisaqueous buffer was added to the flask containing the coating of lecithinand cholesterol, and this mixture was stirred for 3-4 hours. During thistime, the coating of lecithin and cholesterol was dispersed in solution.A cloudy solution resulted.

[0187] Emulsion

[0188] In a 400 mL, thick-walled beaker, were mixed highly refined acidfree olive oil 31.49 g, and oleic acid 3.51 g. The emulsifier solutiondescribed above was then added to bring the total weight of the mixtureto 350 g. A 1-inch drying coated stir bar was added, and the mixture wasstirred magnetically for 2-5 minutes. The mixture was then irradiatedwith 2 bursts of ultrasound (45 sec. each, with 2 minutes of magneticstirring between bursts) using a Branson Sonifier 450 operated atmaximum power with a ¾″ solid horn. The pH of the resulting emulsion wasadjusted to 6.5 (at 20° C.). The emulsion prepared in this way was usedimmediately in the fat binding test, but could be kept in a refrigerator(4° C.) for a week.

[0189] When the physiological emulsion described above was mixed withtest polymers, it was observed that a solid polymer/lipid complex wouldform in some cases. A test was devised to measure the quantity of lipidabsorbed by the test polymers from the physiological emulsion.

[0190] Lipid Binding Test

[0191] The test polymer (25 mg) was weighed into a tared 20 mLcentrifuge filter cup with a 10 micron polypropylene mesh filter(Whatman VECTASPIN20™ centrifuge filter). The bottom of the filter cupwas then sealed with tape to prevent solution from leaking out duringthe test. Using an analytical pipette, an aqueous buffer solution (3 mL)containing NaCl (0.1M), and MES (0.1M) at pH=6.5 was added to the filtercup. The filter cup was inserted into its companion centrifuge tube andsealed with a cap. This assembly was agitated in an orbital shaker forat least 1 hour in order to dissolve or disperse the test polymer. Theolive oil emulsion described above (15 mL) was then added to the filtercup using an analytical pipette. The cap was replaced, and thecentrifuge tube shaken (250 rpm) on an orbital mixer for a period of onehour. The centrifuge filter device was then disassembled so that thetape could be removed from the bottom of the centrifuge filter cup. Itwas immediately reassembled and spun in a centrifuge at an RCF of 500 G,and at 25° C. for 30 minutes. The centrifuge filter device was removedfrom the centrifuge and disassembled. The filter cup was weighed toobtain the weight gain of the wet polymer/lipid complex. This materialwas the removed from the filter cup with a spatula, and placed into atared glass vial. The vial was weighed again to obtain the weight of thepolymer/lipid sample. The vial was then placed into a centrifugalevaporator, and dried at 60° C. under vacuum until a pressure of 0.15Torr or less was achieved (8-18 hrs). The vial was removed and weighedto obtain the dry weight of the polymer/lipid complex sample. The amountof lipid absorbed by the original 25 mg polymer sample in the filter cupwas then calculated. This gravimetric result was used as a measure forlipid binding by the polymer, and is listed in the accompanying table aslipid weight absorbed (g) per gram of polymer. TABLE 15 Lipid weight (g)absorbed EXAMPLE No. by 1 gram of polymer Chitosan 2.2 10 5.5 13 6.8 1420 15 44.9 16 3.4 52 3.3 54 4.6 55 3.8 66 5.9 67 5.9 69 4.5 70 4.1 725.8 75 4.5 76 3.9 78 3 79 3.6 80 12.6 81 3.1 83 10 84 2.7 85 11.3 86 2.187 6.5 88 3.1 89 6.9 90 4.5 91 6.2 92 10.9 94 2.8 96 4.7 97 4.5 98 1.799 5.7 100 3.3 101 11 102 13.4 103 4.3 104 16 105 2.1 107 7.7 108 6.6109 5.1 110 6.1 111 4.8 112 3.8 113 60 114 58 115 58 116 59 117 59 11860 119 54 120 59 121 60 122 56 123 54 124 50 125 59 126 52 127 56 128 66129 62 130 60 131 59 132 58 133 59 134 49 135 49 136 43 137 57 138 64140 63 141 66 166 48 167 59 168 61 169 39 172 61 173 33 174 39 175 65176 15 177 37 178 21 179 24 180 57 181 68 182 60 183 51 184 65 185 7 18667 187 27 188 2 189 43 190 10 191 2 192 68 193 67 194 61 195 64 196 16197 8 198 13 199 5 202 9 203 6 204 15 205 7 206 3 207 22 208 4 209 3

[0192] In vivo Testing of Fat-Binding Polymers

[0193] The non-crosslinked and crosslinked fat-binding polymers ofExamples 5, 6, 10, 72, 173 and Chitosan were evaluated for their abilityto increase the excretion of fat in the feces, relative to the controlgroup, in normal rats over a six-day period. Male Sprague-Dawley rats(five to six weeks of age) were individually housed and fed ad libitum apowdered “high-fat diet,” consisting of standard rodent chowsupplemented with 15% lard by weight. After feeding the animals thisdiet for five days, the animals were weighed and sorted into thetreatment or control groups (4-6 animals per group, each group havingequal mean body weights). Animals were treated for six days with thetest compounds, which were added to the “high-fat diet” atconcentrations (w/w) of 0.0% (control), 2.0 or 5.0 percent of the diet.In one study chitosan was evaluated for its effect on fecal fatexcretion.

[0194] Rat fecal samples were collected on the final three days of thesix days of drug treatment. The samples were freeze dried and ground toa fine powder. One half gram of sample was weighed and transferred toextraction cells. Samples were extracted in an accelerated solventextractor (ASE 200 Accelerated Solvent Extractor, Dyonex Corporation,Sunnyvale, Calif.) with 95% ethanol, 5% water and 100 mM KOH. The samplewas extracted in 17 minutes at 150° C. and 1500 psi. An aliquot ofextract was transferred to a test tube containing a molar excess of HCl.The sample was then evaporated and reconstituted in a detergent solutionconsisting of 2% Triton X-1200, 1% polyoxyethylene lauryl ether and 0.9%NaCl. Fatty acids were then quantitated enzymatically with acolorimetric kit (NEFAC, Wako Chemical GmbH, Neuss, Germany).

[0195] Table 16 contains values for fecal fat excretion as a percentageof ingested fat. TABLE 16 IN VIVO EFFICACY OF FAT-BINDING POLYMERS FECALFAT Dose EXCRETION % (w/w percent OF INGESTED Example Identification ofdiet) FAT Example 6-PAA 3% XL, 5 40 10% C12 Example 6-PAA 3% XL, 10% C122 12 Example 5-PAA 9.4% XL 2 22 Example 10-PDA 4.5% XL 2 30 Example72-PDA 4.5% XL, 11% C12 2 29 Example 173-TMAEMC (90 mol %) + 2 20octadecyl methacrylate(10 mol %) Chitosan 2  8

[0196] Fecal Fat/Consumed Fat was calculated as follows: Fatty acidconcentration from the enzymatic assay was expressed as mmol/mL. Themmol/mL of fatty acid was then multiplied by the number of mL of extractgenerated from 500 mg of sample to give the total mmol of fatty acid.The value for the total mmol of fatty acid was converted to total mg offatty acid using the average molecular weight of medium to long chainfatty acid (270 D). The value was corrected for any dilutions madeduring sample workup. When results are expressed as mg/gm of feces, thetotal mg of fatty acids is multiplied by 2. When results were expressedas total mg of fatty acid excreted in 24 hours, the mg/gm of feces valuewas multiplied by fecal weight in grams excreted in 24 hours. When theresults were expressed as excreted fat as a % of that consumed in 24hours, the total weight of fat excreted in 24 hours was divided by theweight of fatty acids consumed over 24 hours and multiplied by 100.

[0197] While this invention has been particularly shown and describedwith references to preferred embodiments thereof, it will be understoodby those skilled in the art that various changes in form and details maybe made therein without departing from the spirit and scope of theinvention as defined by the appended claims. Those skilled in the artwill recognize or be able to ascertain using no more than routineexperimentation, many equivalents to the specific embodiments of theinvention described specifically herein. Such equivalents are intendedto be encompassed in the scope of the claims.

What is claimed is:
 1. A method for treating obesity in a mammal,comprising the step of orally administering to the mammal an effectiveamount of one or more fat-binding polymers in combination with one ormore lipase inhibitors. 2 The method of claim 1 wherein the lipaseinhibitor is selected from lipstatin, tetrahydrolipstatin or acombination thereof.
 3. The method of claim 1 wherein the fat-bindingpolymer comprises a monomer of the formula

wherein R is a hydrophobic region.
 4. The method of claim 3 herein thehydrophobic region comprises a substituted or unsubstituted, normal,branched or cyclic alkyl group having at least about four carbons. 5.The method of claim 3 wherein the hydrophobic region comprises an alkylgroup of between about four and thirty carbons. 6 The method of claim 3wherein the hydrophobic region comprises an alkyl group of about 6carbons.
 7. A method for reducing the absorption of dietary fat in amammal, comprising the step of orally administering to the mammal atherapeutic amount of one or more fat-binding polymers in combinationwith one or more lipase inhibitors.